Tag: GFP

. 2023 May 9;14(22):5945-5955. doi: 10.1039/d3sc00629h. eCollection 2023 Jun 7. Affiliations Expand Affiliations 1 Beijing Key Laboratory for Bioengineering and Sensing Technology, School of Chemistry and Biological Engineering, University of Science and Technology Beijing 30 Xueyuan Road, Haidian District Beijing 100083 P. R. China. 2 Department of Chemistry, Tsinghua University…

Cell lines and culture A549-hACE2 [a gift from Dr. Ben tenOever3], 293 T, HEK293, A549, and 3T3-L1 pre-adipocyte cell lines (refer to Supplementary Table 1) were cultured in DMEM and 10% FBS. U937, Calu-3, and HCT116 cells were cultured in RPMI, EMEM, and McCoy’s 5 A media, respectively, with 10% FBS. Caco-2 cells…

Ethics statement All animal studies were performed in accordance with the recommendations in the Canadian Council on Animal Care Guidelines. The reporting of this data in the manuscript follows the recommendations in the ARRIVE guidelines. The University of Calgary Health Sciences Animal Care Committee approved all animal protocols and surgical…

Isolation of amino acid auxotrophic mutants using the CRISPR/Cas9 system To allow pDUAL-type single crossover chromosomal integration, strains having a point mutation in genes preferably encoding proteins functioning in the amino acid or nucleotide biosynthesis pathway are needed. There are several such auxotrophic mutants in fission yeast. However, the mutation…

Cell culture and cell line construction Cells were cultured at 37 °C in 5% CO2 atmosphere with 100% humidity. Telomerase-immortalized RPE-1 retinal pigment epithelium cells (CRL-4000, American Type Culture Collection), U2OS osteosarcoma cells (HTB-96, American Type Culture Collection) and derivative cell lines were grown in DMEM/F12 (1:1) medium without phenol red…

Expression and purification of human PTH1R The plasmid encoding human PTH1R (GenBank identifier: U17418.1; residues 27–491) was constructed and purified as previously reported17. The construct was expressed in HEK293 GnTI (N-acetylglucosaminyltransferase I-negative) cells (American Type Culture Collection, CRL-3022) using the BacMam system (Thermo Fisher Scientific), and the cells were grown…

NOTCH3 anti-peptide antibodies NOTCH3 is a transmembrane receptor (Fig. 1A) that plays multiple roles in development, homeostasis, and pathology. Mutations in NOTCH3 are responsible for the most common cause of inherited stroke and vascular dementia, cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL)8,9. Figure 1 Generation and characterization of…

Re: [PATCH 1/5] uio: Add hv_vmbus_client driver – kernel test robot From: kernel test robot <lkp@intel.com> To: Saurabh Sengar <ssengar@linux.microsoft.com>, kys@microsoft.com, haiyangz@microsoft.com, wei.liu@kernel.org, decui@microsoft.com, mikelley@microsoft.com, gregkh@linuxfoundation.org, linux-kernel@vger.kernel.org, linux-hyperv@vger.kernel.org Cc: oe-kbuild-all@lists.linux.dev Subject: Re: [PATCH 1/5] uio: Add hv_vmbus_client driver Date: Sun, 4 Jun 2023 10:05:10 +0800 [thread overview] Message-ID: <202306040910.13oLufQd-lkp@intel.com> (raw)…

Insertion of nucleic acids into cells unlocks the possibility of modulating gene expression; however, some cells such as primary cells and those that grow in suspension are hard-to-transfect. New therapies for cancer and possible autoimmune diseases, such as those that involve chimeric antigen receptor T cells, rely on the insertion…

We present a protocol for assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq) specifically on adipocytes using nucleus sorting with adipose tissues isolated from transgenic reporter mice with nuclear fluorescence labeling. ATAC-seq is a powerful technique to understand gene expression regulatory mechanisms. However, adipose tissue is difficult using this technique…

DOI: 10.1016/j.vaccine.2007.10.041 CAS-3 JCR-Q3 SCIE Lauren A. HiraoLing WuAmir S. KhanAbhishek SatishchandranRuxandra Draghia-AkliDavid B. Weiner Lauren A. HiraoLing WuAmir S. KhanAbhishek SatishchandranRuxandra Draghia-AkliDavid B. Weiner 摘要 In an effort to improve DNA vaccine immune potency electroporation has emerged as a method of delivery of plasmids to target tissues. However, few…

Hi all, I am expressing a GFP synonymous variant library in human cells and sequencing its RNA on the nanopore and I am having some trouble analysing the data. Initially, I basecalled all the fast5 files using the super accuracy model in the guppy basecaller, then I discarded the reads…

Phytopathogenic bacterial cytidine base editor system. a A four-module system to make gene specific cytidine base editor. Expression cassettes of guide RNA (gRNA), SacB, and chimeric gene of dead Cas9 (dCas9), cytidine deaminase (CDA1) and DNA uracil glycosylase inhibitor (UGI) each are flanked by the Gateway recombination motifs (attR and…

Assembly of T4 artificial viral vectors T4-AVVs were assembled by sequential incorporation of purified biomaterials to generate a virus structural mimic (Fig. 2a and Supplementary Movie 1). Starting with an empty capsid shell purified from E. coli infected by the neck-minus and tail-minus T4 phage mutant (10-amber.13-amber.HocΔ.SocΔ T4) (Supplementary Fig. 1a), a pentameric…

Statement on ethical considerations All animal work was approved and permitted by the Local Ethical Committee on Animal Experiments and conducted according to the Guidelines for Animal Experimentation recommendations (ARRIVE guidelines). In particular, mouse work related to C57BL/6NCrl mice was approved and permitted by the Institute of Molecular Genetics of…

Vector construction The Cas9/sgRNA vector (all-in-one) (GenBank accession no. OQ579018) contained sgRNA and Cas9-2A-mCherry expression cassettes. The sgRNA (5′-ATGCAGAACTAGAGTACAGC-3′) targets the phiC31 pseudo-attP intergenic site located on chromosome 3 of CHO-K1 genome (GenBank accession no. APMK01032147.1) (Fig. 1a). To construct the Cas9-mSA/sgRNA vector (GenBank accession no. OQ579019), the monomeric streptavidin (mSA)…

Appl Microbiol Biotechnol . 2023 May 26. doi: 10.1007/s00253-023-12462-y. Online ahead of print. Affiliations Expand Affiliations 1 Department of Chemical Engineering, University of Waterloo, 200 University Ave. W., Waterloo, N2L 3G1, Ontario, Canada. 2 Department of Chemical Engineering, University of Waterloo, 200 University Ave. W., Waterloo, N2L 3G1, Ontario, Canada….

[PATCH] When ‘mcf_edma’ is allocated, some space is allocated for a flexible array at the end of the struct. ‘chans’ item are allocated, that is to say ‘pdata->dma_channels’. * [PATCH] When ‘mcf_edma’ is allocated, some space is allocated for a flexible array at the end of the struct. ‘chans’ item…

CRISPR editing of DNA polymerase δ To increase the genetic repertoire of P. falciparum parasites in culture, we aimed to generate parasites with impaired 3ʹ−5ʹ proof-reading activity from the catalytic subunit of DNA polymerase δ (PF3D7_1017000) in order to increase the level of basal spontaneous mutations, based on prior work…

This protocol describes how to use TIRF microscopy to track individual ion channels and determine their activity in supported lipid membranes, thereby defining the interplay between lateral membrane movement and channel function. It describes how to prepare the membranes, record the data, and analyze the results. Ion channels are not…

[PATCH 1/3] w1: gpio: Don’t use platform data for driver data – Uwe Kleine-König From: “Uwe Kleine-König” <u.kleine-koenig@pengutronix.de> To: Krzysztof Kozlowski <krzysztof.kozlowski@linaro.org> Cc: Rob Herring <robh@kernel.org>, Greg Kroah-Hartman <gregkh@linuxfoundation.org>, linux-kernel@vger.kernel.org, Philipp Marek <philipp@marek.priv.at> Subject: [PATCH 1/3] w1: gpio: Don’t use platform data for driver data Date: Thu, 25 May 2023…

[PATCH 3/3] w1: gpio: rename pointer to driver data from pdata to ddata – Uwe Kleine-König From: “Uwe Kleine-König” <u.kleine-koenig@pengutronix.de> To: Krzysztof Kozlowski <krzysztof.kozlowski@linaro.org> Cc: Rob Herring <robh@kernel.org>, Greg Kroah-Hartman <gregkh@linuxfoundation.org>, linux-kernel@vger.kernel.org, Philipp Marek <philipp@marek.priv.at> Subject: [PATCH 3/3] w1: gpio: rename pointer to driver data from pdata to ddata Date:…

Mutant collection development We mutagenized 2,700 seeds of the wheat–Th. elongatum introgression line RWG34 containing Sr43 (ref. 29). Dry seeds were incubated for 16 h with 200 ml of a 0.8% (w/v) EMS solution with constant shaking on a Roller Mixer (Model SRT1, Stuart Scientific) to ensure maximum homogenous exposure of the…

Catalog number MNSI000A-1 Price Please ask Size 10 ug Shipping Temperature Blue Ice/ Dry Ice Product category Minicircle Technology Description Isotype or positive controls by peptides, antibodies and deactivated samples. Kit Plasmid mini made and maxi DNA purification kits can be silica gel or anion exchange, endotoxin free and are…

Author Contributions Conceptualization, L.W., W.B. and V.G.; methodology, Y.H., J.S. (John Sienkiewicz), J.S. (Jinying Sun), H.Z., H.H., S.X., R.B. and L.H.; software, Y.H., J.S. (John Sienkiewicz) and J.S. (Jinying Sun); validation, H.Z. and J.S. (John Sienkiewicz); formal analysis, V.G.; investigation, V.G. and L.W.; resources, L.W.; data curation, Y.H., J.S. (John…

Introduction Microalgae encompass a large number of organisms with both prokaryotic and eukaryotic nature, making them an archetypal platform for recombinant technology (Malla et al., 2021). The nutraceutical significance of microalgae is characterized by their rich natural biomolecules and high amount of protein, vitamins, and lipid substances (Potvin and Zhang, 2010;…

Molecular profiling reveals common changes in matched pairs of HNSCC primary tumor and metastasis-derived cell lines Identification of both genetic and functional differences between primary and metastatic variants of HNSCC would be enabled by in vitro models derived independently from these sites from the same patient. Unfortunately, HNSCC tumor cells…

Catalog No. PVTBAV11741-1 Vector Backbone pLenti-shRNA-GFP-Puro Backbone manufacturer N/A Vector type Lentiviral interference Backbone size w/o insert 11.6kb Cloning site 5 N/A Site destroyed during cloning N/A Cloning site 3 N/A Bacterial resistance Ampicillin Growth strain Stbl3 Growth temperature 37 Centigrade High or low copy High Copy Selectable markers Puromycin…

A. Microarray data (GSE12777) for 51 breast cancer cell line was analyzed using Gene-E software. E-cadherin correlated 121 genes are enriched in first step followed by EpCAM close (correlated). These genes are presented in normal and Ras-transduced cells (right panel). B-C. Two independent microarray data sets from MCF10A and Ras-transduced…

Clinical spectrum The clinical findings of the 15 individuals are summarized in Table 1. Comparison of the clinical presentation of this cohort with previously reported cases is summarized in Table 2. Pedigrees are shown in Fig. 1a and detailed clinical reports are provided in Supplementary File 2 and Supplementary Figure 1. Table 1 Clinical information….

Modification of HDRTs with interstrand crosslinks increases HR during gene editing. (a) Top panel: Cas9 RNPs introduce a double strand DNA break (DSB) at a targeted region in the genome, which can be repaired by error prone end joining (EJ) processes that rejoin the ends of the break, or homology-directed…

Cells and cell culture KOPN30, BV173, and K562 are BCR/ABL1-positive leukemia cell lines. All leukemia cell lines, as well as Ba/F3 cells, were maintained in RPMI-1640 medium supplemented with 15% fetal bovine serum (FBS) and penicillin–streptomycin (100 U/mL) at 37 °C in an atmosphere containing 5% CO2. KOPN30 cells were obtained…

. 2023 Apr 20;14:1179394. doi: 10.3389/fpls.2023.1179394. eCollection 2023. Affiliations Expand Affiliation 1 College of Forestry, Fujian Agriculture and Forestry University, Fuzhou, China. Item in Clipboard Shanwen Ye et al. Front Plant Sci. 2023. Show details Display options Display options Format AbstractPubMedPMID . 2023 Apr 20;14:1179394. doi: 10.3389/fpls.2023.1179394. eCollection 2023. Affiliation 1…

1 // SPDX-License-Identifier: GPL-2.0-only 2 /* 3 * Toshiba TC6387XB support 4 * Copyright (c) 2005 Ian Molton 5 * 6 * This file contains TC6387XB base support. 7 */ 8 9 #include <linux/module.h> 10 #include <linux/platform_device.h> 11 #include <linux/clk.h> 12 #include <linux/err.h> 13 #include <linux/mfd/core.h> 14 #include <linux/mfd/tmio.h> 15…

Imagine you a working on a new CRISPR class 2 endonuclease called Cas99. You have already shown that it can be “programmable” against GFP (green fluorescent protein) in bacterial cells by adding a single guide RNA called sgRNA-GFP that matches a 20 bp stretch of the GFP sequence, adjacent to…

Ethics statement All experiments in this study comply with guidelines of the U.S. Public Health Service and were approved by the Institutional Biosafety Committee at the University of Pittsburgh. All animal husbandry and experimental procedures were conducted in accordance with U.S. Public Health Service policy and approved by the Institutional…

Here, we present a protocol to isolate murine intestinal mesenchyme, including telocytes. These can be used for several applications, such as co-culture with mouse or human-derived organoids, to support growth and better reflect the situation in the original tissue. Our protocol for isolating intestinal mesenchyme results in a high yield…

Ethics declaration Animal study was performed following the Guide for the Care and Use of Laboratory Animals. Animal experiment was approved by the Institutional Animal Care and Use Committee of Colorado State University (protocol number 1090) in advance and conducted in compliance with the Association for the Assessment and Accreditation…

The RNA-guided nucleases derived from the CRISPR-Cas systems in bacteria and archaea have found numerous applications in biotechnology, with genome redaction, imaging, plus gene regular. Does, an discovery of novel Cas nucleases holds outpaced yours characterization and succeeding exploitation. A key step in characterizing Cas nucleases is determining which protospacer-adjacent…

CLIP enables high-efficiency knock-in into an essential locus To create CLIP, we leverage integrase-deficient lentiviruses (IDLV) to deliver an HDR donor template as a viral RNA genome to cells25,26. IDLV consists of lentiviral components but with a D64V mutation in the integrase, maintaining this enzyme’s ability to package and nuclear…

Transcribed image text: 3. You now plan to clone the PCR amplicon for the WT FANCG gene into the PvuII site of plasmid pGreen as illustrated below. The PCR product is a linear, blunt-ended dsDNA molecule of 360 bp, with an AfaI cleavage site at position 354. pGreen is 2000bp…

Transcriptional activation of RCOR2 by neo-TADs in RELA ependymoma. a Chromatin contacts in a reconstructed ZFTA tumor genome (sample 4EP53) including the tandem duplication that leads to the ZFTA-RELA fusion (chr1:63532174-65429788, green boxes). Solid black boxes show TADs identified by applying TopDom to the Hi-C data mapped on to the…

All methods were performed in accordance with institutional guidelines and regulations. CRISPR-Cas9 protein and guide RNA reagents All Cas9 proteins used in this study were derived from the prototypic Streptococcus pyogenes species. Cas9 protein was produced as described in Seamon et al.33 and contained a N-terminal nuclear localization signal and…

Delivery of mRNA to Jurkat and primary pan T cells To demonstrate the capability of our platform to deliver mRNA, we transfected Jurkat and primary T cells with mRNA encoding GFP. Primary T cells from healthy donors were transfected 4 days after activation with CD3/CD28 antibodies. Both cell types were resuspended…

SARS-CoV-2 infection induces mtDNA release and recognition by cGAS Recent studies revealed that SARS-CoV-2 stimulation could cause the accumulation of intracellular and intercellular DNA due to cell fusion, cell death and even tissue damage.17,19,27 To detect SARS-CoV-2-triggered release of DNA during infection, we first detected the cytosolic dsDNA pattern by…

Expression of PPP2R3B/ppp2r3b in human foetuses and zebrafish To begin to investigate the requirement for PR70 in vertebrates, we looked for expression of PPP2R3B in human foetuses. The orientation of tissue sections used is given in Supplementary Fig. 1. Using in situ hybridisation, we noted some locations of PPP2R3B expression that…

[PATCH v6 07/13] coresight: Store pointers to connections rather than an array of them – James Clark From: James Clark <james.clark@arm.com> To: coresight@lists.linaro.org, quic_jinlmao@quicinc.com, mike.leach@linaro.org, suzuki.poulose@arm.com Cc: James Clark <james.clark@arm.com>, Leo Yan <leo.yan@linaro.org>, Alexander Shishkin <alexander.shishkin@linux.intel.com>, Mathieu Poirier <mathieu.poirier@linaro.org>, Greg Kroah-Hartman <gregkh@linuxfoundation.org>, linux-arm-kernel@lists.infradead.org, linux-kernel@vger.kernel.org Subject: [PATCH v6 07/13] coresight: Store…

Bba_K4501010 Device rgasra Pau Contains exosome booster genes such as STEAP3, SDC4 and NadB and a puromycin resistance gene for selection. BBa_K4501012 Composite Exosome booster 2 Eva Contains exosome booster gene nSMase and a hygromycin resistance gene for selection. BBa_K4501013 Composite CD63_His-tag Jaume Contains the tetraspannin CD63 labelled with 6x…

Abstract All 81 ribosomal proteins (RPs) that form the Arabidopsis (Arabidopsis thaliana) 80S ribosome are encoded by several paralogous genes. For example, the nearly identical RPS24A and RPS24B proteins are encoded by RPS24A and RPS24B, respectively. Here we explored the functions of RPS24A and RPS24B in Arabidopsis. Their encoding genes…

Cell lines and tissue culture The murine 17-CL1 cell line (derived from 3T3 cells, BEI Resources, Cat. No. NR-53719) was maintained in Dulbecco Modified Eagle Medium (DMEM, Life Technologies) supplemented with 10% Fetal Bovine Serum (FBS, Gemini Bio-Products), L-glutamine (Invitrogen), and Sodium pyruvate (Invitrogen). The NCTC clone 1469 cell line…

Hello Kah Jing Lee, The patch 1b4394c5d731: “firmware: stratix10-svc: extend svc to support RSU feature” from Jul 11, 2022, leads to the following Smatch static checker warning: drivers/firmware/stratix10-svc.c:627 svc_normal_to_secure_thread() warn: ‘pdata->command == 10’ ‘false’ implies ‘pdata->command != 10’ is ‘true’ drivers/firmware/stratix10-svc.c 383 static int svc_normal_to_secure_thread(void *data) 384 { 385 struct…

[PATCH 1/2] dt-bindings: regulator: Add dt property * [PATCH 1/2] dt-bindings: regulator: Add dt property @ 2023-04-18 14:50 Naresh Solanki 2023-04-18 14:50 ` [PATCH 2/2] regulator: userspace-consumer: Multiple regulators Naresh Solanki 2023-04-20 0:01 ` [PATCH 1/2] dt-bindings: regulator: Add dt property Zev Weiss 0 siblings, 2 replies; 4+ messages in…

Physicochemical properties of siRNA-encapsulated LNPs and their endocytosis. (A) Structure of the ionizable lipid 10 used for LNP preparation. (B) Composition of the particles. (C) Average size (in nm), zeta potential (in mV), PDI (polydispersity index), and % encapsulation efficiency of the particles. Data are represented as mean ± SEM…

Small interfering RNAs (siRNAs) are novel therapeutics that can be used to treat a wide range of diseases. This has led to a growing demand for selective, efficient, and safe ways of delivering siRNA in cells.  Now, in a cooperation between the universities of Amsterdam and Leiden in the Netherlands,…

Cell culture and reagents HEK293 and HeLa cells (American Type Culture Collection) were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal calf serum, 2 mM GlutaMAX‐1, 100 μg/mL streptomycin and 100 U/mL penicillin (complete growth medium) at 37 °C under 5% CO2. HeLa cells stably expressing GFP-TIFA were previously described8. Cells…

  WATERTOWN, Mass., April 18, 2023 /PRNewswire/ — Cultivarium, the focused research organization creating open-source tools to expand access to uncharacterized microorganisms, and ATCC, the premier global biological materials management and standards organization, have entered a multi-phase research collaboration to expand the use of non-model microorganisms. The…

Abstract African Swine Fever (ASF) disease is caused by infection of susceptible Suidae with African Swine Fever Virus (ASFV). In Eurasian domestic pigs, and closely related wild boar, ASF is characterised by a haemorrhagic fever and a high mortality rate of 95-100%. There are currently no effective treatments for ASF,…

Transcribed image text: Exercise 3 The following assay looks at the role of Cas 9 and tracrRNA in regulation of the Stroptococcus Pyogenes CRISPR/Cas system. For this, an in vitro experiment was designed in which only a few components are used. The mothod for this assay is called Electromobily shift…

Genome-scale CRISPR screen to identify regulators that maintain mESC pluripotency To establish a function-based PGRN, we first performed a CRISPR-Cas9 mediated genome-wide screen to detect genes essential for self-renewal. mESCs were cultured under Leukaemia inhibitory factors (LIF)/serum condition (L/S), which was commonly used in similar tasks and confer a naïve…

Figure 1. Schematic of Cre reporter design. GFP, pA, and FLuc indicate green fluorescent protein, terminator polyadenylation signal, and firefly luciferase, respectively. Before recombination, the reporter gene is transcribed to synthesize GFP (black arrow) but not FLuc (X). Cre protein catalyzes the recombination between the two loxP sites flanking the…

Search for creTA elements associating with I-F CRISPR-Cas Previously identified creTA genes all surround the cas6 gene of a type I or III CRISPR-Cas19. Yet, within the I-F CRISPR-Cas loci of Acinetobacter species, we did not find any creTA-like elements by searching the sequences surrounding csy4 (a subtype-specific cas6 gene)….

SygRNA modified synthetic tracrRNA and crRNAs are optimized for use with a variety of Cas9 expressing cell lines, Cas9 proteins, or Cas9 mRNAs and are …SygRNA® synthetic Francisella novicida tracrRNA complexed with SygRNA® synthetic Francisella novicida crRNA. For use with MISSION® FnCas9 proteins in …16 nov. 2016 — SygRNA™-Synthetic Two Part CRISPR…

Abstract Cationic polymers are potential intracellular carriers for small interfering RNA (siRNA). The short and rigid nature of an siRNA chain often results in larger and more loosely packed particles compared to plasmid DNA (pDNA) after complexing with carrier polycations, and in turn, poor silencing effects are seen against the…

Cell culture We cultured mESCs in t2iL medium containing Dulbecco’s modified eagle medium (DMEM, Nacalai Tesque), 2 mM Glutamax (Nacalai Tesque), 1× non-essential amino acids (Nacalai Tesque), 1 mM sodium pyruvate (Nacalai Tesque), 100 U ml−1 penicillin, 100 μg ml−1 streptomycin (P/S) (Nacalai Tesque), 0.1 mM 2-mercaptoethanol (Sigma) and 15% fetal bovine serum (FBS) (Gibco), supplemented with…

Transcriptional dysregulation has been implicated in a number of progressive neurodegenerative diseases. To correct transcriptional misregulation, emerging CRISPR tools such as CRISPR activation (CRISPRa) and CRISPR interference (CRISPRi) could be used. In addition, efficient and precise transcriptional activation in broad brain areas and multiple neuroanatomical structures can be achieved by…

OTUB1 deficiency causes defects in osteogenesis To determine the physiological role of OTUB1 in bone homeostasis, Otub1 global knockout mice (referred to as Otub1−/−) were constructed, and their phenotypes were analyzed on embryonic (E) day 14.5 (E14.5), E16.5, E18.5, and postnatal (P) day P0 (Fig. 1a and Supplementary Fig. 1a,…

BsAb structural and compositional considerations, and combinatorial library design The structural and compositional attributes of BsAb design have a significant impact on their functionalities. Unlike the trial-and-error approach used in conventional methods, we can conduct unbiased interrogation of many structural and compositional variables (e.g., affinity, epitope location, linker design, and…

Introduction Among all common noncutaneous malignancies, bladder carcinoma (BC) is the fourth prevalent one in all patients from the United States, and a subset of BC can progress to a severe muscle invasive form, leading to distal metastasis to other organs, including liver, lung, bone and mediastinum (1). BC with…

For this case study, we selected seven protein sequences for common biotechnology tools, ranging in length from 18 to 231 amino acids. One of these is the T4 foldon, a short sequence from an E. coli bacteriphage that has a long history of use for stabilization of proteins (e.g.10,11), including…

Cell culture HEK293T, MEF-1, and U87 cell lines were used in this research. HEK293T was purchased from ATCC. MEF-1, and U87 cell lines were gifts from Muredach P. Reilly, Peter A. Sims, respectively. HEK293T, MEF-1, and U87 cells were cultured in DMEM with 4.5 g/L D-Glucose, supplemented with 10% fetal bovine…

[PATCH v3 02/13] coresight: Change name of pdata->conns – James Clark From: James Clark <james.clark@arm.com> To: coresight@lists.linaro.org, quic_jinlmao@quicinc.com, mike.leach@linaro.org, suzuki.poulose@arm.com Cc: James Clark <james.clark@arm.com>, Mathieu Poirier <mathieu.poirier@linaro.org>, Leo Yan <leo.yan@linaro.org>, Alexander Shishkin <alexander.shishkin@linux.intel.com>, Maxime Coquelin <mcoquelin.stm32@gmail.com>, Alexandre Torgue <alexandre.torgue@foss.st.com>, linux-arm-kernel@lists.infradead.org, linux-kernel@vger.kernel.org, linux-stm32@st-md-mailman.stormreply.com Subject: [PATCH v3 02/13] coresight: Change name of…

Affiliations Expand Affiliations 1 Laboratory of Experimental Virology, Department of Medical Microbiology, Amsterdam UMC, Academic Medical Center, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands. 2 Department of Molecular and Cell Biology, National Center of Biotechnology (CNB-CSIC), Campus Universidad Autónoma de Madrid, 28049 Madrid, Spain. Free PMC article Item in…

CRISPR RNA is delivered to the mouse lung by inhalation using improved lipid nanoparticles. Many patients with inherited lung diseases lack effective treatment and could derive immense benefit from gene-editing therapies that correct the causative mutation. Direct delivery of such therapies to the lung by inhalation would be transformative, but…

(A) Immunofluorescence images of spinal cord sections from wild-type Δ7SMA mice at 25 weeks that received AAV9-ABE + AAV9-GFP in a 10:1 ratio by neonatal ICV injection, stained for GFP to indicate AAV transduction, NeuN as a marker of post-mitotic neurons, and DAPI to stain all nuclei. (B) In vivo…

Introduction Bacteriophage (phage) is a class of widespread viruses exclusively infecting bacteria, playing a key role in ecosystems.1–3 Since phages were discovered over a hundred years ago, many techniques and reagents, such as DNA polymerase, restriction endonuclease, ligase, and CRISPR/Cas system, have been developed from phages, which have together accelerated…

Plasmid construction The PVCpnf structural and accessory region (pvc1-16) and payload and regulatory region (Pdp1, Pnf and regulatory genes PAU_RS16570-RS24015) were synthesized de novo (GenScript) and cloned into pAWP78 and pBR322 backbones, respectively. All manipulations involving payload and regulatory plasmids (pPayload) involved standard PCR amplification with Phusion Flash 2x Master…

Published: 28 March 2023| Version 1 | DOI: 10.17632/fwczkb6xw3.1 Contributors: Description This study was designed to capture the changes in CD8 T cell phenotypes in murine glioma (CT-2A) model post immunotherapy with AAV-LIGHT (TNFSF14). Tumour infiltrating CD45 cells were isolated by flow sorting and subject to targeted single-cell transcriptome sequencing…

Research reported in this study complies with all relevant ethical regulations, and our animal study protocol has been approved by the Animal Experimental Board of Finland (license number ESAVI/28687/2020. Phage panning Sherpabody phage display library (size ~1011 cfu) was obtained from Next Biomed Therapies Oy. To develop sherpabodies specific for SARS-CoV-2…

Mice Rosa26-YFP48, Rosa-tDTomato49, K14creER, Lgr5creER (ref. 50), KrasLSLG12D (ref. 51) and Trp53fl/fl (ref. 52) mice were obtained from the NCI mouse repository and Jackson Laboratories. Rhojfl/fl mice53 were a gift from A. Uemura. All mice used in this study were composed of males and females with mixed genetic background. Mouse…

The CHO cell line is an adherent epithelial cell derived from the ovary of Chinese hamster. It is widely used in biology and medical research of genetics, toxicity screening, nutrition, and gene expression, particularly to express recombinant proteins. CHO cells are the most common mammalian hosts for industrial production of…

Patient samples and RNA-seq analysis RNA-seq data from 395 patients with PDA was obtained from the University Health Network (Toronto), Sunnybrook Health Sciences Centre (Toronto), Kingston General Hospital (Kingston), McGill University (Montreal), Mayo Clinic (Rochester), Massachusetts General Hospital (Boston) and Sheba Medical Center (Tel Aviv) and has been described previously1,12,13,14….

To examine the function of the Scribble polarity aenetwo shRNA seauences(shScrib1 and shScrib2) have been designed to knockdown Scribble expression. To test their effectiveness,along with a negative control(empty vector)and a positive control(shGFP)(which can also knockdown the Scribble-GFP fusion)they have been tested on HEK293 cells that stably express Scribble GFP In…

Experimental animals and DOX treatment All animal experiments performed on live animals were approved by Fudan University’s Committee on the Ethics of Animal Experiments and adhered to relevant guidelines including the ARRIVE guidelines for animal experiments in the study. Adult male C57BL/6 mice (8–10 weeks old; 23–26 g; Department of Laboratory…

Yarmolinsky, D. A., Zuker, C. S. & Ryba, N. J. Common sense about taste: from mammals to insects. Cell 139, 234–244 (2009). Article  CAS  PubMed  PubMed Central  Google Scholar  Liman, E. R., Zhang, Y. V. & Montell, C. Peripheral coding of taste. Neuron 81, 984–1000 (2014). Article  CAS  PubMed  PubMed…

. 2023 Mar 16;18(3):e0282689. doi: 10.1371/journal.pone.0282689. eCollection 2023. Affiliations Expand Affiliations 1 Center of Life Sciences, Skolkovo Institute of Science and Technology, Moscow, Russia. 2 Peoples’ Friendship University of Russia (RUDN University), Moscow, Russia. 3 Armand Hammer United World College of the American West, Montezuma, New Mexico, United Stated of…

I was trained in cell biology and biophysics and, as I was often reminded by study section reviewers, I am “not a cancer biologist”. My closest collaborator, the cell biologist Gunter Blobel6, with whom I had worked for 25 years, encouraged me to leap off in this new direction, building upon…

Transcribed image text: A. What type of FACS data graphical representations are Panel A and Panel B? (1 Mark) B. All data displayed in Panel B have been gated from data displayed in Panel A using a FSC and SSC gate. What does this statement mean? Why would you want…

With an easy-to-use instrument to encapsulate single cells in picolitre-volume water droplets (picodroplets) at high throughput rates of thousands of droplets per second, picodroplet microfluidics is quickly emerging as the technology of choice for single-cell analysis. This highly effective and delicate method has important benefits in terms of cost and…

SygRNA Cas9 Synthetic Modified tracrRNA; HPLC-purified SygRNA modified tracrRNA oligo is optimized for use with a target-specific crRNA or modified crRNA …The SygRNA™ synthetic RNA system recruits SpCas9 nuclease to specific genomic targets. The tracrRNA (trans-activating crRNA) binds to a crRNA (CRISPR RNA) …16 nov. 2016 — SygRNA™-Synthetic Two Part CRISPR RNA…

Ethics declarations This study was approved by the Animal Care and Use Committee of Tottori University (Permit Number: 16-Y-20, 17-Y-28, 19-Y-22, 20-Y-31, 21-Y-26, and 22-Y-36). All experiments were carried out in compliance with the ARRIVE guidelines. All methods were performed in accordance with the relevant guidelines and regulations. Mice were…

Screening for hyper-potent shRNA against SARS-CoV-2 We parsed the SARS-CoV-2 genome into a series of potential short hairpin RNA (shRNA) targets (Supplemental Fig. 1). This process was conducted by tiling the genome with overlapping 50 nucleotide-long sequences, each shifted by a single nucleotide from the other. The region targeted by each…

Chemicals Quinine hydrochloride and denatonium benzoate were obtained from Fujifilm Wako Pure Chemical (Osaka, Japan) and dissolved in a buffer or normal water. Collagenase A and dispase II were from Roche (Basel, Switzerland). Elastase was from Worthington Biochemical (Lakewood, NJ, USA). DNase I was from Merck (Darmstadt, Germany). Lipofectamine 2000…

Animals All the procedures and protocols were approved by the Institutional Animal Care and Use Committee of the National Institute for Basic Biology (15A005, 14A003, 13A023, 12A020, 11A028) and Kyushu University (A21-043-0, A19-137-0, A19-137-1, A19-137-2, A29-088-0, A29-088-1, A29-088-2). Japanese medaka (Oryzias latipes) were bred and maintained under artificial reproductive conditions…

I’ve been trying to run a minimal example of the NextFlow RNAseq pipeline, like so: nextflow run nf-core/rnaseq -r 3.10.0 -profile test,docker –outdir /home/kai/RNASeq/rnaseq_test/test_output However, this appears to return the error below: Error executing process > ‘NFCORE_RNASEQ:RNASEQ:PREPARE_GENOME:GTF2BED (genome_gfp.gtf)’ Caused by: Process `NFCORE_RNASEQ:RNASEQ:PREPARE_GENOME:GTF2BED (genome_gfp.gtf)` terminated with an error exit status (1)…

Mice All mice experiments were performed in accordance with the guidelines of the Institutional Animal Care and Use Committee (IACUC) at the Center for Excellence in Molecular Cell Science, Shanghai Institutes of Biological Sciences, Chinese Academy of Science. The approved animal protocol number is SIBCB-S374-1702-001-C1. The CK19-CreER, HNF4α-DreER, R26-tdTomato (R26-tdT),…

General Questions What information should I know before I do the RIP-Seq? Before doing a RIP-SEQ you generally need to specify • The type of sample to be done • What RBPs are of interest • Whether IP-level antibodies are available for these RBPs • The type…

Collection and maintenance of Drosophila biauraria We used laboratory stocks of Drosophila biauraria (Diptera; Drosophilidae), which were originally collected at the Field Science Center for Northern Biosphere, Hokkaido University located at Tomakomai, Hokkaido in 2011 and 2015 using standard banana traps and sweeping22. Females were brought into the lab and…

Date Sat, 11 Mar 2023 19:28:05 +0000 From Jonathan Cameron <> Subject Re: [PATCH v1 2/4] misc: adps990x: convert to OF On Wed, 8 Mar 2023 11:02:17 +0200Svyatoslav Ryhel <clamor95@gmail.com> wrote:> Add ability to get essential values from device tree.> > Signed-off-by: Svyatoslav Ryhel <clamor95@gmail.com>Hi,Some comments inline.> —> drivers/misc/apds990x.c |…

   Image transcription textcompterERCREDriverSTOPDriverNature Reviews | NephrologyFigure 3 | Schematic of the Cre-fox system used to mark pericytes.a | Transgenic mouse “A” contains Cre recombinase capable ofrecognizing fox sites, under the control of a specific promoter (inthis case Foxd), a transcription factor that is expressed indeveloping mesenchyme cells and later…

[PATCH v2 5/9] coresight: Dynamically add connections – James Clark From: James Clark <james.clark@arm.com> To: coresight@lists.linaro.org Cc: James Clark <james.clark@arm.com>, Mathieu Poirier <mathieu.poirier@linaro.org>, Suzuki K Poulose <suzuki.poulose@arm.com>, Mike Leach <mike.leach@linaro.org>, Leo Yan <leo.yan@linaro.org>, Alexander Shishkin <alexander.shishkin@linux.intel.com>, linux-arm-kernel@lists.infradead.org, linux-kernel@vger.kernel.org Subject: [PATCH v2 5/9] coresight: Dynamically add connections Date: Fri, 10 Mar…