Tag: EDTA

A Method to Obtain CTCs from Murine Colorectal Cancer Model

– Circulating Tumor Cells, or CTCs, are the cancer cells that stem from the primary tumor and enter the blood circulation. Most CTCs can survive in the bloodstream without attaching to the surface; therefore, we can detect them directly from the blood of most cancer patients. CTCs in the blood…

Continue Reading A Method to Obtain CTCs from Murine Colorectal Cancer Model

96 | ELISA Kit for Mitochondrial Open Reading Frame Of The

Specificity This assay has high sensitivity and excellent specificity for detection of Mitochondrial Open Reading Frame Of The 12S rRNA-c (MOTS-c).No significant cross-reactivity or interference between Mitochondrial Open Reading Frame Of The 12S rRNA-c (MOTS-c) and analogues was observed. Recovery Matrices listed below were spiked with certain level of recombinant…

Continue Reading 96 | ELISA Kit for Mitochondrial Open Reading Frame Of The

Metagenomic analysis of Mesolithic chewed pitch reveals poor oral health among stone age individuals

The specific environmental/history/collection context The Huseby Klev materials were unearthed and collected by archaeologists (including two of the co-authors of this article) during the excavation of this coastal hunter-fisher-gatherer site in the 90s50. The material assemblage was rich and well preserved: human bones, animal bones, plant remains and pieces of…

Continue Reading Metagenomic analysis of Mesolithic chewed pitch reveals poor oral health among stone age individuals

Detection of DNA methylation signatures through the lens of genomic imprinting

Animals and samples The study included 10 pigs, 8 pigs were bred at the INRAE experimental farm (https://doi.org/https://doi.org/10.15454/1.5572415481185847E12) and 2 pigs come from breeding organizations in accordance with the French and European legislation on animal welfare. The animals belong to the same family, except for one LW animal. Animals were…

Continue Reading Detection of DNA methylation signatures through the lens of genomic imprinting

Proinflammatory chemokine CXCL14 activates MAS-related G protein-coupled receptor MRGPRX2 and its putative mouse ortholog MRGPRB2

Reagents and peptides CXC motif chemokine 14 (CXCL14) was purchased from BIOZOL (Eching, Germany). Proadrenomedullin N-terminal 20 peptide (PAMP-20) was obtained from Tocris Bioscience (Bristol, UK), and cortistatin-14 was from GenScript (NJ, USA). Custom synthesis of CXCL14 analogs was performed by JPT Peptide Technologies (Berlin, Germany) (see Supplementary Fig. 6). Stock…

Continue Reading Proinflammatory chemokine CXCL14 activates MAS-related G protein-coupled receptor MRGPRX2 and its putative mouse ortholog MRGPRB2

Domestic pigs are susceptible to experimental infection with non-human primate-derived Reston virus without the need for adaptation

Ethics and animal welfare statement All infectious work with RESTV, including sample inactivation, was performed in the Containment Level 4 laboratory (CL4) in accordance with the policies and protocols outlined by the Canadian Science Centre for Human and Animal Health Institutional Biosafety Committee. All animal work was performed in strict…

Continue Reading Domestic pigs are susceptible to experimental infection with non-human primate-derived Reston virus without the need for adaptation

ACADVL Recombinant Protein | Scientist.com

ACADVL Recombinant Protein | Scientist.com Skip to Main Content Go to Main Navigation Details Alternative Name: ACAD6||LCACD||VLCAD. Amino Acid: MGSSHHHHHH SSGLVPRGSH MAGGAAQLAL DKSDSHPSDA LTRKKPAKAE SKSFAVGMFK GQLTTDQVFP YPSVLNEEQT QFLKELVEPV SRFFEEVNDP AKNDALEMVE ETTWQGLKEL GAFGLQVPSE LGGVGLCNTQ YARLVEIVGM HDLGVGITLG AHQSIGFKGI LLFGTKAQKE KYLPKLASGE TVAAFCLTEP SSGSDAASIR TSAVPSPCGK YYTLNGSKLW ISNGGLADIF TVFAKTPVTD PATGAVKEKI TAFVVERGFG GITHGPPEKK MGIKASNTAE VFFDGVRVPS ENVLGEVGSG…

Continue Reading ACADVL Recombinant Protein | Scientist.com

Apoptosis-resistant megakaryocytes produce large and hyperreactive platelets in response to radiation injury | Military Medical Research

Animals C57BL/6-Tg (Pf4-icre) Q3Rsko/J (Pf4-Cre) mice were purchased from the Jackson Laboratory (Bar Harbor, ME, USA). B6;129-Tmem173tm1(flox)Smoc (Stingfl) mice were purchased from Shanghai Model Organisms Center, Inc. (Shanghai, China). C57BL/6J-Ifnar1em1(flox)Cya (Ifnar1fl) mice were purchased from Cyagen Biosciences (Guangzhou, China). Stingcko and Ifnar1cko mice were generated by crossing Pf4-Cre mice with…

Continue Reading Apoptosis-resistant megakaryocytes produce large and hyperreactive platelets in response to radiation injury | Military Medical Research

Analysis of off-tumour toxicities of T-cell-engaging bispecific antibodies via donor-matched intestinal organoids and tumouroids

Human tissue samples Tissue samples and annotated data were obtained, and experimental procedures were performed within the framework of the non-profit foundation HTCR, including informed patient consent. Intestinal organoid and tumouroid cell culture Supplementary Table 1 provides anonymized basic donor information. After isolation of intestinal crypts following a previously described…

Continue Reading Analysis of off-tumour toxicities of T-cell-engaging bispecific antibodies via donor-matched intestinal organoids and tumouroids

Preclinical assessment of an anti-HTLV-1 heterologous DNA/MVA vaccine protocol expressing a multiepitope HBZ protein | Virology Journal

Design of the recombinant virus MVA-HBZ The multiepitope protein design was based on HBZ sequence deposited in the GenBank database [17, 23, 24], and in silico analyses performed using the epitope prediction tools: NetMHCpan 4.0 (Threshold: weak binders—2.0; strong binders—0.5), NetCTL 1.2 (Threshold of 0.75) and IEDB—MHC-I Binding Predictions. Epitopes…

Continue Reading Preclinical assessment of an anti-HTLV-1 heterologous DNA/MVA vaccine protocol expressing a multiepitope HBZ protein | Virology Journal

Genomic hypomethylation in cell-free DNA predicts responses to checkpoint blockade in lung and breast cancer

Lung cancer ICB cohort Advanced non-small cell lung carcinoma patients who were treated with anti-PD-1/PD-L1 monotherapy at Samsung Medical Center, Seoul, Republic of Korea were enrolled for this study. The present study has been reviewed and approved by the Institutional Review Board (IRB) of the Samsung Medical Center (IRB no….

Continue Reading Genomic hypomethylation in cell-free DNA predicts responses to checkpoint blockade in lung and breast cancer

Gadolinium Oxide Nanoparticles reinforce immune response

Boyi Yu,1– 4 Xuanyi Lu,5 Xianglong Feng,1– 4 Ting Zhao,1– 4 Jiaxin Li,1– 4 Yudie Lu,5 Fei Ye,1– 4 Xiongxiong Liu,1– 4 Xiaogang Zheng,1– 4 Zheyu Shen,5 Xiaodong Jin,1– 4 Weiqiang Chen,1– 4 Qiang Li1– 4 1Biomedical Center, Institute of Modern Physics, Chinese Academy of Sciences, Lanzhou, People’s Republic of…

Continue Reading Gadolinium Oxide Nanoparticles reinforce immune response

Conserved and divergent gene regulatory programs of the mammalian neocortex

Nucleus preparation from frozen brain tissue for Chromium single-cell multiome ATAC and gene expression analysis M1 tissue was obtained from three human donors (male, aged 42, 29 and 58 years), three macaque donors (male, aged 6 (Macaca mulatta), 6 (M. mulatta) and 14 (Macaca fascicularis) years), three marmoset (Callithrix jacchus)…

Continue Reading Conserved and divergent gene regulatory programs of the mammalian neocortex

Bioactive glycans in a microbiome-directed food for children with malnutrition

Collection and handling of biospecimens obtained from participants in the randomized controlled clinical study of the efficacy of MDCF-2 The human study entitled ‘Community-based clinical trial with microbiota-directed complementary foods (MDCFs) made of locally available food ingredients for the management of children with primary moderate acute malnutrition (MAM)’ was approved…

Continue Reading Bioactive glycans in a microbiome-directed food for children with malnutrition

The landscape of genomic structural variation in Indigenous Australians

Cohorts Saliva and/or blood samples were collected from consenting individuals among four NCIG-partnered communities: Tiwi Islands (comprising the Wurrumiyanga, Pirlangimpi and Millikapiti communities), Galiwin’ku, Titjikala and Yarrabah, between 2015 and 2019. Non-Indigenous comparison data, generated from unrelated Australian individuals of European ancestry, was drawn from two existing biomedical research cohorts:…

Continue Reading The landscape of genomic structural variation in Indigenous Australians

Regulation of the ER-Resident Mannosidase EDEM2 in HEK293 Cells

Abstract EDEM2 plays an important role as the first enzyme that acts during mannose trimming of N-glycosylated proteins in the ERAD machinery. Although EDEM2 expression has been reported to be transcriptionally regulated by the IRE1-sXBP1 pathway, very little is known about how endogenous EDEM2 protein expression is regulated. In this…

Continue Reading Regulation of the ER-Resident Mannosidase EDEM2 in HEK293 Cells

Human RAD52 stimulates the RAD51-mediated homology search

Introduction Homologous recombination (HR) is an evolutionarily conserved process that plays a pivotal role in genome stability, diversity, and plasticity. HR is indeed a key repair pathway able to faithfully repair DNA damages including double-strand breaks (DSBs) and DNA gaps by copying the error-free information from the template DNA normally…

Continue Reading Human RAD52 stimulates the RAD51-mediated homology search

Thyroid hormone-regulated chromatin landscape and transcriptional sensitivity of the pituitary gland

Mouse genetic models The ThrbHAB allele expresses TRβ proteins (TRβ1 and TRβ2) fused to a peptide with a hemagglutinin (HAx2) tag and a site for biotinylation by prokaryotic BirA ligase, modified from a published tag30. The tag was inserted at the endogenous Thrb gene by homologous recombination in W9.5 (129/Sv)…

Continue Reading Thyroid hormone-regulated chromatin landscape and transcriptional sensitivity of the pituitary gland

Biosensors | Free Full-Text | CRISPR/Cas12a-Based Detection Platform for Early and Rapid Diagnosis of Scrub Typhus

1. Introduction Orientia tsutsugamushi (OT) is an obligate intracellular parasite bacteria and the causative agent of scrub typhus (ST), which is associated with acute febrile illness (AFI) [1] and transmitted by mites through an infected chigger bite (in the larval stage). This disease, which was earlier believed to be endemic…

Continue Reading Biosensors | Free Full-Text | CRISPR/Cas12a-Based Detection Platform for Early and Rapid Diagnosis of Scrub Typhus

Carbohydrates and carbohydrate degradation gene abundance and transcription in Atlantic waters of the Arctic

Seawater samples were collected from surface waters (SRF) and the bottom of the surface mixed layer (BML) in the Eastern Fram Strait region to investigate the distribution of carbohydrates and their utilisation by microbial communities. The ten sites were grouped into three categories based on the underlying seafloor topography (above-slope,…

Continue Reading Carbohydrates and carbohydrate degradation gene abundance and transcription in Atlantic waters of the Arctic

Generating high-quality plant and fish reference genomes from field-collected specimens by optimizing preservation

Sample collection A total of nine species of marine fish were collected across three different sampling days (September 7th, 9th, and 12th 2022) under IACUC Animal Use Protocol S12219 (Supplementary Data 1). Six species were collected using a speargun donated by a local fisher. Fish were transported back to shore, euthanized,…

Continue Reading Generating high-quality plant and fish reference genomes from field-collected specimens by optimizing preservation

DNA polymerases in precise and predictable CRISPR/Cas9-mediated chromosomal rearrangements | BMC Biology

Cell culture The human endometrial carcinoma HEC-1-B cells were cultured in the modified Eagle’s medium (MEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin at 37°C in a 5% (v/v) CO2 incubator. The human embryonic kidney HEK293T cells were cultured in the Dulbecco’s modified Eagle’s medium (DMEM) supplemented…

Continue Reading DNA polymerases in precise and predictable CRISPR/Cas9-mediated chromosomal rearrangements | BMC Biology

Endometrial cancer PDX-derived organoids (PDXOs) and PDXs with FGFR2c isoform expression are sensitive to FGFR inhibition

Patients, tumour Implantation, PDX expansion and characterization The study was conducted according to the Declaration of Helsinki. Tumours were collected from EC patients following Human Research Ethics Committee (HREC) approval (HREC/15/MHS/127) and QUT HREC (#1500000169, 1500000323). All participants provided written informed consent that their excess tissue could be used for…

Continue Reading Endometrial cancer PDX-derived organoids (PDXOs) and PDXs with FGFR2c isoform expression are sensitive to FGFR inhibition

Phenotypic profiling of solute carriers characterizes serine transport in cancer

Cell culture All cell lines used in this study were cultured at 37 °C in 5% CO2 in a humidified incubator. Human cell lines were authenticated by STR profiling using Promega GenePrint 10 and tested for Mycoplasma using Mycoalert (Lonza). Other than HCT116 p21−/− (a gift of B. Vogelstein60) all cell…

Continue Reading Phenotypic profiling of solute carriers characterizes serine transport in cancer

Gene association analysis of an osteopontin polymorphism and ketosis resistance in dairy cattle

Methods Approval for animal experiments The prof. Dorota Ziyba-Przybylska as a Chairman of ethics review board and Dean of the Faculty of Animal Science, University Agriculture in Krakow considers that this type of project does not fall under the legislation for the protection of animals used for scientific purposes. The…

Continue Reading Gene association analysis of an osteopontin polymorphism and ketosis resistance in dairy cattle

Genomic DNA extraction optimization and validation for genome sequencing using the marine gastropod Kellet’s whelk [PeerJ]

Introduction Rapidly advancing next generation sequencing technologies such as whole genome sequencing, genotyping-in-thousands by sequencing (GT-seq), and restriction-site-associated DNA sequencing (RAD-seq), are becoming more available and affordable for non-model organisms (Park & Kim, 2016; Ellegren, 2014; Van Wyngaarden et al., 2017; Bootsma et al., 2020). However, applying these technologies to…

Continue Reading Genomic DNA extraction optimization and validation for genome sequencing using the marine gastropod Kellet’s whelk [PeerJ]

500U | Taq DNA Polymerase, 5u/ul Anawa

Taq DNA Polymerase, 5u/ul Product information Taq DNA Polymerase (New Buffer!) Product Description: Taq DNA Polymerase is a thermosle DNA Polymerase isolated from a strain of Thermus sp. Taq has a half life of 3 hours at 95°C. Taq has high fidelity with an error frequency 10/10^6 (or 0.01/10^3) during…

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Common analysis of direct RNA sequencinG CUrrently leads to misidentification of m5C at GCU motifs

Introduction Oxford Nanopore Technologies (ONT) direct RNA sequencing (Fig 1A) enables detection of RNA modifications. A modified base produces an altered electrical current and/or dwell time relative to a canonical base that can be detected with algorithms (Garalde et al, 2018; Smith et al, 2019; Workman et al, 2019). Figure…

Continue Reading Common analysis of direct RNA sequencinG CUrrently leads to misidentification of m5C at GCU motifs

A Reliable Method for Quantifying Plasma Cell-Free DNA Using an Internal Standard Strategy: Evaluation in a Cohort of Non-Pregnant and Pregnant Women

Martignano F. Cell-free DNA: an overview of sample types and isolation procedures. In: Casadio V, Salvi S, editors. Cell-free DNA as diagnostic markers. Methods in molecular biology, Vol. 1909. New York, NY: Humana Press; 2019. p. 13–27. https://doi.org/10.1007/978-1-4939-8973-7_2. Volckmar AL, Sültmann H, Riediger A, Fioretos T, Schirmacher P, Endris V,…

Continue Reading A Reliable Method for Quantifying Plasma Cell-Free DNA Using an Internal Standard Strategy: Evaluation in a Cohort of Non-Pregnant and Pregnant Women

Two mitochondrial HMG-box proteins, Cim1 and Abf2, antagonistically regulate mtDNA copy number in Saccharomyces cerevisiae | Nucleic Acids Research

Abstract The mitochondrial genome, mtDNA, is present in multiple copies in cells and encodes essential subunits of oxidative phosphorylation complexes. mtDNA levels have to change in response to metabolic demands and copy number alterations are implicated in various diseases. The mitochondrial HMG-box proteins Abf2 in yeast and TFAM in mammals…

Continue Reading Two mitochondrial HMG-box proteins, Cim1 and Abf2, antagonistically regulate mtDNA copy number in Saccharomyces cerevisiae | Nucleic Acids Research

MSL2 ensures biallelic gene expression in mammals

Materials Animals All of the mice were kept in the animal facility of the Max Planck Institute of Immunobiology and Epigenetics. The mice were maintained under specific-pathogen-free conditions, with 2 to 5 mice housed in individually ventilated cages (Techniplast). The cages were equipped with bedding material, nesting material, a paper…

Continue Reading MSL2 ensures biallelic gene expression in mammals

ESRP1 controls biogenesis and function of a large abundant multiexon circRNA | Nucleic Acids Research

Abstract While the majority of circRNAs are formed from infrequent back-splicing of exons from protein coding genes, some can be produced at quite high level and in a regulated manner. We describe the regulation, biogenesis and function of circDOCK1(2–27), a large, abundant circular RNA that is highly regulated during epithelial-mesenchymal…

Continue Reading ESRP1 controls biogenesis and function of a large abundant multiexon circRNA | Nucleic Acids Research

Population-specific distribution of TPMT deficiency variants

Introduction Thiopurine S-methyltransferase (TPMT) is a cytoplasmic enzyme that catalyzes the S-methylation of purine analogs, including azathioprine, 6-mercaptopurine (6-MP), and thioguanine.1 The metabolism of these drugs results in two types of metabolites: S-methylmercaptopurine and S-methylthioguanine, which are generally described as inactive metabolites, and S-methyl-thioinosine monophosphate, an inhibitor of de novo…

Continue Reading Population-specific distribution of TPMT deficiency variants

Optimized RT-qPCR and a novel normalization method for validating circulating miRNA biomarkers in ageing-related diseases

Reagents miRNeasy Serum/Plasma Advanced Kit (Qiagen, Hilden, Germany, # 217204); TaqMan® Advanced miRNA cDNA Synthesis Kit (Applied Biosystems, Bedford, MA, USA, #A28007); TaqMan® Fast Advanced Master Mix (Applied Biosystems, Bedford, MA, USA, # 4444556); TaqMan® Advanced miRNA Assays Single-tube assays (Applied Biosystems, Bedford, MA, USA, # A25576: 478293_mir, Spike-In cel-miR-39-3p;…

Continue Reading Optimized RT-qPCR and a novel normalization method for validating circulating miRNA biomarkers in ageing-related diseases

Biophysical properties of NaV1.5 channels from atrial-like and ventricular-like cardiomyocytes derived from human induced pluripotent stem cells

hiPSC cultures and cardiomyocyte differentiation The hiPSC lines CBRCULi001-A54 and CBRCULi008-A55 were generated from a 44-year-old male and 75-year-old female control lymphoblastoids, respectively, and they were reprogramed at the LOEX core facility (Quebec City, QC, Canada). All the work with hiPSCs were approved by CIUSSS de la Capitale-Nationale ethics committee (Project…

Continue Reading Biophysical properties of NaV1.5 channels from atrial-like and ventricular-like cardiomyocytes derived from human induced pluripotent stem cells

Immune-privileged tissues formed from immunologically cloaked mouse embryonic stem cells survive long term in allogeneic hosts

Mice C57BL/6N (strain 005304), C3H/HeJ (strain 000659), FVB/NJ (strain 001800), BALB/cJ (strain 000651) and NSG mice (stock 005557) were purchased from the Jackson Laboratory. CD-1 (stock 022) mice were purchased from Charles River. Mice (6–20-week-old) of each strain/background were used for teratoma assays. Mice were housed in a pathogen-free facility…

Continue Reading Immune-privileged tissues formed from immunologically cloaked mouse embryonic stem cells survive long term in allogeneic hosts

High-throughput screening of genetic and cellular drivers of syncytium formation induced by the spike protein of SARS-CoV-2

Plasmid construction All the constructs used in this study were generated with standard cloning strategies, including PCR, overlapping PCR, oligo annealing, digestion and ligation. Primers were purchased from Genewiz. The plasmid sequence was verified by Sanger sequencing. The pCAG-spike(D614G)-GFP11-mCherry plasmid was modified from Addgene plasmid 158761. Briefly, GFP11 and mCherry…

Continue Reading High-throughput screening of genetic and cellular drivers of syncytium formation induced by the spike protein of SARS-CoV-2

Cell-free DNA extraction from urine of lung cancer patients and healthy individuals: Evaluation of a simple method using sample volume up-scaling

Background: Urine holds promise as a source for cell-free DNA (cfDNA) analysis of cancer genetics due to its nature as a self-collectable biospecimen available in large quantities. However, pre-analytical variables such as preservation of cfDNA or efficiency of up-scaling specimen volume need to be better explored to increase analysis sensitivity….

Continue Reading Cell-free DNA extraction from urine of lung cancer patients and healthy individuals: Evaluation of a simple method using sample volume up-scaling

The Mla system of diderm Firmicute Veillonella parvula reveals an ancestral transenvelope bridge for phospholipid trafficking

Bacterial strains and growth conditions Veillonella parvula SKV38 was grown in SK medium (10 g/L tryptone [Difco], 10 g/L yeast extract [Difco], 0.4 g/L disodium phosphate, 2 g/L sodium chloride, and 10 ml/L 60% [wt/vol] sodium DL-lactate; described in ref. 51. Cultures were incubated at 37 °C in anaerobic conditions, either in anaerobic bags (GENbag anaero;…

Continue Reading The Mla system of diderm Firmicute Veillonella parvula reveals an ancestral transenvelope bridge for phospholipid trafficking

Megakaryocytes possess a STING pathway that is transferred to platelets to potentiate activation

Introduction Host defence against infection relies on two important systems: the immune and the coagulation systems. Platelets are anucleate cells produced by megakaryocytes and are responsible for blood clotting and haemostasis (Lefrancais et al, 2017). They are also very abundant and play unexpected roles in immune responses (Maouia et al,…

Continue Reading Megakaryocytes possess a STING pathway that is transferred to platelets to potentiate activation

Elevated stress response marks deeply quiescent reserve cells of gastric chief cells

Generation of inducible H2b-GFP knock-in mice Generation of inducible H2b-GFP knock-in mice was performed by using CRISPR/Cas943. In brief, a mixture containing a sgRosa26-1 crRNA43 (8.7 ng/μl, Fasmac, Japan), a tracrRNA (14.3 ng/μl, Fasmac, Japan), a single strand oligo donor nucleotide (ssODN) composed of 5′ arm, adenovirus splicing acceptor, SV40 pA, TRE3G…

Continue Reading Elevated stress response marks deeply quiescent reserve cells of gastric chief cells

Intrinsic deletion at 10q23.31, including the PTEN gene locus, is aggravated upon CRISPR-Cas9-mediated genome engineering in HAP1 cells mimicking cancer profiles

Introduction The CRISPR-Cas system is a widely used genome engineering technology because of its simple programmability, versatile scalability, and targeting efficiency (Wang & Doudna, 2023). Although researchers are rapidly developing CRISPR-Cas9 tools, the biggest challenge remains to overcome undesired on- and off-targeting outcomes. Previous studies have reported unintended genomic alterations,…

Continue Reading Intrinsic deletion at 10q23.31, including the PTEN gene locus, is aggravated upon CRISPR-Cas9-mediated genome engineering in HAP1 cells mimicking cancer profiles

LncRNA INHEG promotes glioma stem cell maintenance and tumorigenicity through regulating rRNA 2’-O-methylation

Ethics statement All mice procedures in this study were performed under an animal protocol approved by the Institutional Animal Care and Use Committee guidelines of Westlake University. The procedures and protocols for glioma patients were approved by the institutional review board of Beijing Tiantan Hospital. Informed consent was obtained from…

Continue Reading LncRNA INHEG promotes glioma stem cell maintenance and tumorigenicity through regulating rRNA 2’-O-methylation

GDF11 slows excitatory neuronal senescence and brain ageing by repressing p21

Experimental model and subject details Mice Male ICR mice (Laboratory Animal Center of Zhejiang Academy of Medical Sciences) at age of 3 months (M), 9 M and 36 M, male C57BL/B6 wild‐type (WT) mice (Shanghai Slac Laboratory) at age of 3 M and 10 M, male GDF11-flox mice (GDF11f/f, mice carrying the “floxed” GDF11…

Continue Reading GDF11 slows excitatory neuronal senescence and brain ageing by repressing p21

IDG-SW3 Cell Culture in a Three-Dimensional Extracellular Matrix

Here, we present a protocol for culturing IDG-SW3 cells in a three-dimensional (3D) extracellular matrix. We detailed a protocol to establish a 3D system using an extracellular metrics along with collagen one for analyzing formation and gene expression through osteogenesis. Extracellular metrics can increase the transparency of the collagen one…

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Primate-specific ZNF808 is essential for pancreatic development in humans

Subjects The study was conducted in accordance with the Declaration of Helsinki and all subjects or their parents/guardian gave informed written consent for genetic testing. DNA testing and storage in the Beta Cell Research Bank was approved by the Wales Research Ethics Committee 5 Bangor (REC 17/WA/0327, IRAS project ID…

Continue Reading Primate-specific ZNF808 is essential for pancreatic development in humans

DRIP-qPCR validation – Mendeley Data

Description 1. HEK293 cells were obtained from the National Collection of Authenticated Cell Culture of China (accession number: SCSP-5014) and cultured in DMEM supplemented with 10% FBS and penicillin/streptomycin (100U/ml) at 37°C with 5%CO2. Expression vector carrying NOTCH2NLC exon1 with different numbers of GGC repeats were prepared and transfected into…

Continue Reading DRIP-qPCR validation – Mendeley Data

PPP1R12A is a recycling endosomal phosphatase that facilitates YAP activation

Antibodies Antibodies used in the present study were as follows: rabbit anti-phospho-YAP (S127) (#13008, dilution 1:500), rabbit anti-phospho-LATS1 (S909) (#9157, dilution 1:500), rabbit anti-YAP (#14074, dilution 1:100 for immunofluorescence), and rabbit anti-PPP1R12A (#8574, dilution 1:1000 for western blot, 1:200 for immunofluorescence) from Cell Signaling Technology; rabbit anti-LATS1 (A300-477A, dilution 1:2000)…

Continue Reading PPP1R12A is a recycling endosomal phosphatase that facilitates YAP activation

Circulating cell-free DNA fragmentation is a stepwise and conserved process linked to apoptosis | BMC Biology

DNA fragmentation of apoptotic cells is a two-step process which gives rise to major and minor peaks at different places DNA ladder formation is the most well-known characteristic of apoptosis. Since the 1990s, the HL60 cell line has been regarded as an ideal model for studying cell apoptosis [15]. We therefore took advantage of…

Continue Reading Circulating cell-free DNA fragmentation is a stepwise and conserved process linked to apoptosis | BMC Biology

A Cre-dependent massively parallel reporter assay allows for cell-type specific assessment of the functional effects of non-coding elements in vivo

Animal models All procedures involving animals were approved by the Institutional Animal Care and Use Committee (IACUC) at Washington University in St. Louis, MO. Veterinary care and housing was provided by the veterinarians and veterinary technicians of Washington University School of Medicine under Dougherty lab’s approved IACUC protocol. All protocols…

Continue Reading A Cre-dependent massively parallel reporter assay allows for cell-type specific assessment of the functional effects of non-coding elements in vivo

Recovery of microbial DNA by agar-containing solution from extremely low-biomass specimens including skin

Ethical considerations This study complies with relevant institutional, national, and international guidelines and legislation. The Ethics Committee at the RIKEN Center for Integrative Medical Sciences (H30-4) approved this study, and written informed consent was obtained from all volunteers who provided specimens. Sampling solutions The AgST solution was prepared as follows:…

Continue Reading Recovery of microbial DNA by agar-containing solution from extremely low-biomass specimens including skin

FAT10 is phosphorylated by IKK{beta} to inhibit the antiviral type-I interferon response

Introduction The innate immune system represents the host´s first-line defense against viral infections (Koyama et al, 2008). In this context, IFN-I are produced by all nucleated cells and represent the principal cytokines that counteract viral replication (Ivashkiv & Donlin, 2014). In fact, several receptors of the infected cell can recognize…

Continue Reading FAT10 is phosphorylated by IKK{beta} to inhibit the antiviral type-I interferon response

Mapping Mammalian 3D Genome Interactions with Micro-C-XL

A protocol for mapping the three-dimensional genome organization with nucleosome resolution using the genome-wide chromosome conformation capture method Micro-C-XL is presented here. This protocol is significant as it shows high resolution chromosome loops and other short-range interaction features. The main advantage of this technique is the mapping of the 3D…

Continue Reading Mapping Mammalian 3D Genome Interactions with Micro-C-XL

Unzipped chromosome-level genomes reveal allopolyploid nematode origin pattern as unreduced gamete hybridization

Nematode materials and species identification Mi, Mj, and Mg were collected from farmlands in Wuhan city of Hubei province, Longyan city of Fujian province, and Changsha city of Hunan Provinces, respectively. Two Ma samples were collected from farmlands in Shenyang city of Liaoning province and Shiping city of Yunnan province….

Continue Reading Unzipped chromosome-level genomes reveal allopolyploid nematode origin pattern as unreduced gamete hybridization

Rapid in situ RNA imaging based on Cas12a thrusting strand displacement reaction | Nucleic Acids Research

Abstract RNA In situ imaging through DNA self-assembly is advantaged in illustrating its structures and functions with high-resolution, while the limited reaction efficiency and time-consuming operation hinder its clinical application. Here, we first proposed a new strand displacement reaction (SDR) model (Cas12a thrusting SDR, CtSDR), in which Cas12a could overcome…

Continue Reading Rapid in situ RNA imaging based on Cas12a thrusting strand displacement reaction | Nucleic Acids Research

An FcRn-targeted mucosal vaccine against SARS-CoV-2 infection and transmission

Mice and Golden Syrian hamsters All research experiments in this study comply with all relevant ethical regulations. The Institutional Animal Care and Use Committee approved the animal protocol at the University of Maryland (#R-APR-20-18 and #R-MAR-21-19). The animals were acclimatized at the animal facility for 4–6 days before initiating experiments….

Continue Reading An FcRn-targeted mucosal vaccine against SARS-CoV-2 infection and transmission

The DNA-binding induced (de)AMPylation activity of a Coxiella burnetii Fic enzyme targets Histone H3

Statistics and reproducibility Anti-AMP IP for LC-MS/MS analysis was performed in three independent biological replicates (n = 3). TSA assay data represents technical triplicates. Time-resolved (de)AMPylation analyzed by LC-MS was performed as biological triplicates. CD measurements were performed in technical triplicates. Anisotropy data is shown as technical triplicates. Anisotropy measurements were repeated…

Continue Reading The DNA-binding induced (de)AMPylation activity of a Coxiella burnetii Fic enzyme targets Histone H3

Highly oxidized albumin is cleared by liver sinusoidal endothelial cells via the receptors stabilin-1 and -2

List of reagents Chloramine-T trihydrate (Merck, Darmstadt, Germany), Copper(II)Sulphate, Penicillin, Streptomycin, RPMI-1640 (Sigma-Aldrich, Burlington, MA, USA), RPMI-1640 (Euroclone, Pero, Italy), DMEM low glucose (Sigma-Aldrich), Trypsin–EDTA (Sigma-Aldrich), Blasticidin hydrochloride (Sigma-Aldrich), Trichloroacetic acid (Merck), Fetal Bovine Serum (Merck, Darmstadt, Germany), Iodine 125 Radionuclide (Perkin Elmer, Waltham, Mass., USA), Iodogen™ iodination reagent (Pierce,…

Continue Reading Highly oxidized albumin is cleared by liver sinusoidal endothelial cells via the receptors stabilin-1 and -2

Cell-Free DNA from Blood or Bone Marrow as Alternatives to Bone Marrow Cells for Molecular Diagnostics and Monitoring of Multiple Myeloma

Oral and Poster Abstracts 652. Multiple Myeloma: Clinical and Epidemiological: Poster III Research, adult, Translational Research, assays, Plasma Cell Disorders, bioinformatics, Diseases, Lymphoid Malignancies, computational biology, Technology and Procedures, Study Population, Human, molecular testing, omics technologies Dor D. Abelman1,2, Jenna Eagles1*, Stephanie Pedersen1*,…

Continue Reading Cell-Free DNA from Blood or Bone Marrow as Alternatives to Bone Marrow Cells for Molecular Diagnostics and Monitoring of Multiple Myeloma

CRISPR/Cas9 bioluminescence-based assay for monitoring CFTR trafficking to the plasma membrane

Introduction Cystic fibrosis (CF) is a lethal, autosomal recessive disorder caused by mutations in the CFTR gene that results in aberrant transport of chloride, bicarbonate, and sodium ions (Kerem et al, 1989; Riordan et al, 1989; Rommens et al, 1989; Tabcharani et al, 1991; Poulsen et al, 1994; Stutts et…

Continue Reading CRISPR/Cas9 bioluminescence-based assay for monitoring CFTR trafficking to the plasma membrane

Comparison of DNA Extraction Methods for the Detection of Canned Tuna Species with DNA Mini-Barcoding

Tuna is susceptible to species mislabeling due to its high demand, quick rate of production, and wide range of price points. DNA barcoding, a sequencing-based technique, allows for the detection of species mislabeling by targeting a standardized region of DNA. A mitochondrial control region (CR) DNA barcode has been found…

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Widespread and largely unknown prophage activity, diversity, and function in two genera of wheat phyllosphere bacteria

Isolation of phyllosphere isolates All bacterial strains were isolated in June 2021 from the flag leaves of four wheat cultivars (Sheriff, Heerup, Rembrandt and Kvium) grown in an experimental field in Høje Taastrup, near Copenhagen, Denmark. Wheat flag leaves were picked, pooled, and either washed or blended prior to dilution…

Continue Reading Widespread and largely unknown prophage activity, diversity, and function in two genera of wheat phyllosphere bacteria

Catalytically inactive long prokaryotic Argonaute systems employ distinct effectors to confer immunity via abortive infection

Bacterial strains and phages Escherichia coli DH5a was routinely grown in Lysogeny broth (LB) medium and used for plasmid cloning, while E. coli BL21 (DE3) was used for protein production and the in vivo assays. E. coli phages T5, T7 and Lambda-vir are gifts from Shi Chen lab (Wuhan University,…

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Synergistic effects of Azithromycin and DMXAA

Kanoktip Petcharat,1 Narongsuk Munkong,2 Rungthip Thongboontho,1 Widsanusan Chartarrayawadee,3 Arthid Thim-Uam1 1Division of Biochemistry, School of Medical Sciences, University of Phayao, Phayao, 56000, Thailand; 2Department of Pathology, School of Medicine, University of Phayao, Phayao, 56000, Thailand; 3Division of Chemistry, School of Science, University of Phayao, Phayao, 56000, Thailand Correspondence: Arthid Thim-Uam,…

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Mouse genome rewriting and tailoring of three important disease loci

BAC plasmids Human (CH17-203N23, CH17-449P15 and CH17-339H2) and mouse (RP23-51O13, RP23-75P20 and RP23-204E8) BACs were purchased from BACPAC Resources Center. Yeast–bacterium shuttle vector pLM1050 was modified by L. Mitchell based on a previous study28. pWZ699 was constructed by inserting a cassette containing pPGK-ΔTK-SV40pA transcription unit and the Actb gene into…

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Circulating DNA and frequency of colorectal cancer brain metastases in a presumed high-risk group

The study was approved by The Central Denmark Region Committees on Health Research Ethics (1-10-72-269-20), and it was prospectively registered with ClinicalTrials.gov (ClinicalTrials.gov identifier: NCT05185557). The research was carried out according to the principles set out in the Declaration of Helsinki 1964 and all subsequent revisions. Written and orally informed…

Continue Reading Circulating DNA and frequency of colorectal cancer brain metastases in a presumed high-risk group

Reduction of retinal ganglion cell death in mouse models of familial dysautonomia using AAV-mediated gene therapy and splicing modulators

Animals All mice were housed in the AALAC-accredited Animal Resource Center at Montana State University. All animal use protocols were approved by the Montana State University Institutional Animal Care and Use Committee (IACUC) (protocol No. 2020-15-IA; Bozeman, MT). The study fulfilled the ARRIVE guidelines, and all experiments complied with relevant…

Continue Reading Reduction of retinal ganglion cell death in mouse models of familial dysautonomia using AAV-mediated gene therapy and splicing modulators

Significant Pathogenic Nontuberculous Mycobacteria | IDR

Introduction The Mycobacterium genus encompasses various species, including the Mycobacterium tuberculosis complex, which is responsible for tuberculosis (TB) infections, Mycobacterium leprae, which associated with leprosy, and a group of nontuberculous mycobacteria (NTM) species.1 The NTM species, also known as Mycobacteria other than tuberculosis (MOTT), can be widely found in the…

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TET2 modulates spatial relocalization of heterochromatin in aged hematopoietic stem and progenitor cells

Animal models Animal studies were approved by the Institutional Animal Care Use Committee of the Institute of Biosciences and Technology, Texas A&M University (AUP 2020-0195). Most mouse strains bear a C57BL/6 genetic background unless otherwise noted. All the animals were housed in a certified animal facility with a standard dark/light…

Continue Reading TET2 modulates spatial relocalization of heterochromatin in aged hematopoietic stem and progenitor cells

Structure and electromechanical coupling of a voltage-gated Na+/H+ exchanger

Expression and purification of SLC9C1 The gene encoding the S. purpuratus SLC9C1 protein (NP_001091927.1), referred to as SLC9C1, was synthesized (Thermo Fisher Scientific) and subcloned into the pcDNA3.1(+) vector followed by a TEV (Tobacco Etch Virus nuclear-inclusion-a endopeptidase) recognition site, eGFP and a TwinStrep tag at the C terminus. An…

Continue Reading Structure and electromechanical coupling of a voltage-gated Na+/H+ exchanger

DNA-binding protein PfAP2-P regulates parasite pathogenesis during malaria parasite blood stages

Parasite culture, maintenance, synchronization and transfection The DiCre-expressing P. falciparum clone II3 (ref. 10) was maintained in human A+ erythrocytes at 37 °C in Roswell Park Memorial Institute 1640 medium containing AlbumaxII (Invitrogen) supplemented with 2 mM l-glutamine. Parasites were either synchronized by sorbitol treatment or by purifying mature schizont stages using…

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Identification of CCZ1 as an essential lysosomal trafficking regulator in Marburg and Ebola virus infections

Cells and viruses Haploid mSCs AN3–12 are a feeder independent clonal derivative of HMSc2 isolated from mice oocyte and maintained at IMBA52. The AN3–12 library and knocked out cells used for the haploid screening was obtained from IMBA (Austria)14,52. AN3–12 cells were validated by STR analysis. Haploid mES cells were…

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Invasive Californian death caps develop mushrooms unisexually and bisexually

Mushroom collecting Sporocarps were collected from various herbaria and during three expeditions to Point Reyes National Seashore (PRNS), California in 2004, 2014 and 2015, and in 2015 from three sites in Portugal. A total of 86 sporocarps were collected: 67 Californian sporocarps (one early herbarium sample dates to 1993), 11…

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PLA2G7 gene polymorphism in gestational diabetes mellitus

Introduction Gestational diabetes mellitus (GDM) is a general term for abnormal glucose tolerance, abnormal fasting blood glucose, and diabetes detected or developed during pregnancy, excluding diabetes that already existed before pregnancy. According to the International Diabetes Federation, the global prevalence of GDM in 2021 was approximately 16.7% and has become…

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Chromatin compartmentalization regulates the response to DNA damage

Cell culture and treatments DIvA (AsiSI-ER-U2OS)19, AID-DIvA (AID-AsiSI-ER-U2OS)23 and 53BP1-GFP DIvA20 cells were developed in U2OS (ATCC HTB-96) cells and were previously described. Authentication of the U2OS cell line was performed by the provider ATCC, which uses morphology and short tandem repeat profiling to confirm the identity of human cell…

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Antiviral type III CRISPR signalling via conjugation of ATP and SAM

Cloning Supplementary Table 1 shows the synthetic gene, DNA and RNA oligonucleotide sequences used in this study. The synthetic genes encoding B.fragilis Cas6, CorA, NrN and C. botulinum SAM lyase purchased as g-blocks (IDT) were codon-optimized for expression in E. coli C43 (DE3) via the vector pEhisV5TEV, which encodes eight…

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Genomic Characteristics of Carbapenem-Resistant Klebsiella pneumoniae

Introduction Klebsiella pneumoniae (K. pneumoniae), which belongs to the family Enterobacteriaceae, is a common nosocomial pathogen that causes a variety of human infections, including bloodstream infection, respiratory tract infection, urinary tract infection, and abdominal infection.1,2 Carbapenems are generally considered to be a last choice for the treatment of multi-drug resistant…

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Plastid-localized xanthorhodopsin increases diatom biomass and ecosystem productivity in iron-limited surface oceans

Protein sequence and prediction analysis The amino acid sequence of FcR was deduced from the F. cylindrus genome sequence33. Protein alignments with characterized microbial rhodopsins were performed using the MUSCLE algorithm implemented in Geneious (v.5.6)69. Predictions of subcellular location were performed with SignalP v.4.0 and TargetP (v.1.1)70. Putative transmembrane domains…

Continue Reading Plastid-localized xanthorhodopsin increases diatom biomass and ecosystem productivity in iron-limited surface oceans

High-resolution analysis of bound Ca2+ in neurons and synapses

Introduction Calcium ions (Ca2+) act as important second messengers in all cells, regulating vital aspects of cellular function from cell proliferation to cell death (Schwarz & Blower, 2016; Giorgi et al, 2018). In the nervous system, Ca2+ signaling plays a special role in the regulation of neuronal and synaptic activities….

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Characterizing the regulatory Fas (CD95) epitope critical for agonist antibody targeting and CAR-T bystander function in ovarian cancer

Mice strains Six- to eight-week-old (age), 20–25-g (weight), both male and female (sex) mice were used for tumor xenografts generation and in vivo efficacy studies as described in the text (see Table 3). Athymic Nude (Envigo) Foxn1nu/Foxn1+ or NOD.Cg Prkdcscid Il2rgtm1Wjl/SzJ also called NSG mice were used. All animal procedures were…

Continue Reading Characterizing the regulatory Fas (CD95) epitope critical for agonist antibody targeting and CAR-T bystander function in ovarian cancer

EOGT enables residual Notch signaling in mouse intestinal cells lacking POFUT1

Single cell RNAseq bioinformatics Bioinformatic trajectory analysis of scRNAseq data obtained from Epcam+CD45− C57Bl6/J adult intestinal crypt cells and deposited as GSE188339 was performed as described35. Primers and antibodies Primer sequences are given in Supplementary Table 1 and antibodies are given in Supplementary Table 2. Generation of Chinese hamster ovary…

Continue Reading EOGT enables residual Notch signaling in mouse intestinal cells lacking POFUT1

Isolation, Culture, and Characterization of Primary Dermal Fibroblasts from Human Keloid Tissue

This study describes an optimized protocol for establishing primary fibroblasts from keloid tissues that can effectively and steadily provide pure and viable fibroblasts. The isolation and culture of primary fibroblasts derived from keloid tissue are the basis for further studies of keloids. Fibroblasts from keloid tissue can be easily acquired…

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Direct regulation of the cardiac ryanodine receptor (RyR2) by O-GlcNAcylation | Cardiovascular Diabetology

Human right atrial appendages (RAA) Human RAA were obtained from consenting patients (ethics: LRS/12/01/001/AM17), with or without Type 2 Diabetic Mellitus (T2DM), undergoing on-pump coronary artery bypass graft surgery. Patient characteristics are shown in Table 1. Patients with pre-operative arrhythmia were excluded from the study. Table 1 Patient characteristics Patients were…

Continue Reading Direct regulation of the cardiac ryanodine receptor (RyR2) by O-GlcNAcylation | Cardiovascular Diabetology

Apoptotic stress causes mtDNA release during senescence and drives the SASP

Cell culture and treatments Human embryonic lung MRC5 fibroblasts (ATCC) and IMR90 fibroblasts (ATCC) were grown in Dulbecco’s modified Eagle’s medium (Sigma-Aldrich, D5796) supplemented with 10% heat-inactivated fetal bovine serum (FBS), 100 U ml−1 penicillin, 100 μg ml−1 streptomycin and 2 mM l-glutamine and maintained at 37 °C under 5% CO2. MRC5 fibroblasts were cultured in…

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A new method for the detection of Mycobacterium tuberculosis based on the CRISPR/Cas system | BMC Infectious Diseases

Expression and purification of Cas12a protein The pMBP-LbCas12a recombinant plasmid used in this study was purchased from Addgene Global Plasmid Information Sharing Platform (Plasmid#113,431) and was 9653 bp in size. The recombinant plasmid was transformed into DH5α competent state and cultured at 37℃ for 6 h. IPTG was added at a final…

Continue Reading A new method for the detection of Mycobacterium tuberculosis based on the CRISPR/Cas system | BMC Infectious Diseases

Genotyping, sequencing and analysis of 140,000 adults from Mexico City

Recruitment of study participants The MCPS was established in the late 1990s following discussions between Mexican scientists at the National Autonomous University of Mexico (UNAM) and British scientists at the University of Oxford about how best to measure the changing health effects of tobacco in Mexico. These discussions evolved into…

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Immunoglobulin superfamily member 3 is required for the vagal neural crest cell migration and enteric neuronal network organization

Immunohistochemistry The tissue sections were processed using the TSA kit (Perkin Elmer) according to the manufacturer’s instructions. The deparaffinization of fixed tissues were performed by immersing the slides in tissue clear and rehydrated with graded ethanol (100–70%) and water. The heat-induced antigen retrieval was performed using citrate buffer (1.8 mM…

Continue Reading Immunoglobulin superfamily member 3 is required for the vagal neural crest cell migration and enteric neuronal network organization

Distribution tendencies of pathogens causing LRTI

Introduction Lower respiratory tract infection (LRTI) remains one of the leading causes of death worldwide.1 Several well-known pathogens, including Streptococcus pneumoniae, Pseudomonas aeruginosa, Klebsiella pneumoniae, Candida, Herpesvirus, and others, have been identified as significant causes of infection.2 Nonetheless, nearly half of the cases still have an undetermined etiology,3,4 despite the…

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Targeted knockout of a conserved plant mitochondrial gene by genome editing

Plant material and growth conditions Nicotiana tabacum cultivar Petit Havana was used for all experiments. The TALEN design and the TALEN-expressing line Nt-JF1006-30 were described previously19. For plant growth under sterile conditions, surface-sterilized seeds were germinated on Murashige and Skoog (MS) medium52 consisting of premixed MS salts and modified vitamins…

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Rapid discovery of high-affinity antibodies via massively parallel sequencing, ribosome display and affinity screening

Construct design To transcribe and translate sequenced DNA clusters on an Illumina flow cell, our DNA constructs contained the following elements: a P5 adaptor, followed by a 28 nt unique barcode, a 27 nt unstructured spacer (5p UNS v2), a ribosome binding site, start codon, protein coding region, TolAk short linker, 2×…

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ncRNA | Free Full-Text | Altered Expression of Vitamin D Metabolism Genes and Circulating MicroRNAs in PBMCs of Patients with Type 1 Diabetes: Their Association with Vitamin D Status and Ongoing Islet Autoimmunity

1. Introduction Type 1 diabetes mellitus (T1DM) is a chronic autoimmune disease characterized by the degeneration of beta cells, resulting in insulin deficiency [1]. T1DM incidence is strongly associated with vitamin D deficiency, and 1,25(OH)2D3 can potentially prevent islet cell death and enhance insulin production. Several studies suggest that low…

Continue Reading ncRNA | Free Full-Text | Altered Expression of Vitamin D Metabolism Genes and Circulating MicroRNAs in PBMCs of Patients with Type 1 Diabetes: Their Association with Vitamin D Status and Ongoing Islet Autoimmunity

Impact of humid climate on rheumatoid arthritis faecal microbiome and metabolites

Animals and husbandry details Six-week-old healthy male SD rats were used in accordance with the Guidelines for the Care and Use of Laboratory Animals of the Institute of Laboratory Animal Resources, Institutional Animal Care and Use Committee of Beijing University of Chinese Medicine. The study is reported in accordance with…

Continue Reading Impact of humid climate on rheumatoid arthritis faecal microbiome and metabolites

Molecular characterization and functional implications on mouse peripheral blood mononuclear cells of annexin proteins from Echinococcus granulosus sensu lato | Parasites & Vectors

Parasites and animals PSCs were collected from hydatid cysts isolated from the liver of sheep infected with E. granulosus s.l. from a slaughterhouse in Sichuan province, China. The PSCs and cyst walls were separated and treated as described in previous reports [22]. The viability and molecular genotyping of PSCs were…

Continue Reading Molecular characterization and functional implications on mouse peripheral blood mononuclear cells of annexin proteins from Echinococcus granulosus sensu lato | Parasites & Vectors

In planta expression of human polyQ-expanded huntingtin fragment reveals mechanisms to prevent disease-related protein aggregation

Plant material and constructs A. thaliana lines of Columbia-0 (Col-0) ecotype were employed, including WT, toc159 (refs. 58,59) and cct8-2 (ref. 60). Seeds underwent surface sterilization and germination on solid 0.5× Murashige and Skoog (M&S) medium with vitamins, lacking sucrose. Plants were incubated in a growth chamber at 22 °C under…

Continue Reading In planta expression of human polyQ-expanded huntingtin fragment reveals mechanisms to prevent disease-related protein aggregation

Multitissue H3K27ac profiling of GTEx samples links epigenomic variation to disease

Samples for H3K27ac ChIP–seq Samples were collected by the GTEx Consortium. The donor enrollment and consent, informed consent approval, histopathological review procedures, and biospecimen procurement methods and fixation were the same as previously described22. No compensation was provided to the families of participants. Massachusetts Institute of Technology Committee on the…

Continue Reading Multitissue H3K27ac profiling of GTEx samples links epigenomic variation to disease

TRIM25 targets p300 for degradation

Introduction Protein levels are regulated at several nodes. One mode of protein level regulation acts through enhancing or reducing gene transcription. Gene transcription can be stimulated by binding transcription factors to promoters or enhancers of target genes and by posttranslational modifications of transcription factors and histones. Such modifications can be…

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Assembloid CRISPR screens reveal impact of disease genes in human neurodevelopment

Culture of hiPS cells The hiPS cell lines used in this study were validated as previously described51. The SEC61B-mEGFP hiPS cell line (AICS-0059 cl.36) was developed at the Allen Institute for Cell Science (https://www.allencell.org/cell-catalog.html) and is distributed through Coriell43. The CAG::Cas9 cell line was genetically engineered in the laboratory (the…

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Schistosoma japonicum EKLF/KLF1 is a potential immune target to tackle schistosomiasis | Parasites & Vectors

Animals and parasites Snails infected with S. japonicum (Oncomelania hupensis) were provided by the Jiangxi Provincial Institute of Parasitic Diseases, Jiangxi, China. Freshly released cercariae stimulated by light were immediately harvested. Six-week-old pathogen-free male BALB/c mice and New Zealand white rabbits (both from Vital River Laboratory Animal Technology Co., Beijing,…

Continue Reading Schistosoma japonicum EKLF/KLF1 is a potential immune target to tackle schistosomiasis | Parasites & Vectors

DNA-bridging by an archaeal histone variant via a unique tetramerisation interface

Chromatin isolation and MNase digestion M. jannaschii DSM 2661 cells were grown in 100 l fermenters in minimal medium containing 0.3 mM K2HPO4, 0.4 mM KH2PO4, 3.6 mM KCl, 0.4 M NaCl, 10 mM NaHCO3, 2.5 mM CaCl2, 38 mM MgCl2, 22 mM NH4Cl, 31 µM Fe(NH4)2(SO4)2, 1 mM C6H9NO6, 1.2 µM MgSO4, 0.4 mM CuSO4, 0.3 µM MnSO4, 36 nM FeSO4, 36 nM CoSO4, 3.5 nM…

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IJNS | Free Full-Text | Whole-Genome Sequencing Can Identify Clinically Relevant Variants from a Single Sub-Punch of a Dried Blood Spot Specimen

1. Introduction Newborn screening (NBS) plays a vital role in healthcare systems for the prompt identification of individuals who may develop one of a set of rare, but severe health conditions [1]. In the UK, for example, dried blood spot (DBS) specimens are routinely collected at 4–5 days of age…

Continue Reading IJNS | Free Full-Text | Whole-Genome Sequencing Can Identify Clinically Relevant Variants from a Single Sub-Punch of a Dried Blood Spot Specimen