Tag: FACS

Unexpected finding in mucus clearance system may inform future pulmonary disease treatment

Generation and validation of MUC5B/AC-KO cultures. (A) Overview of the MUC5B/MUC5AC gene-targeting approach via lentivirus-mediated delivery of sgRNA and CRISPR-Cas9. Transduced and sorted cells were differentiated at air-liquid interface (ALI). Once fully differentiated, the mucus gel was collected by apically washing cultures. HEK293T, human embryonic kidney–293T; FACS, fluorescence-activated cell sorting;…

Continue Reading Unexpected finding in mucus clearance system may inform future pulmonary disease treatment

A potent, broadly protective vaccine against SARS-CoV-2 variants of concern

Cells and viruses hACE2 expressing BHK-21 (BHK-21-hACE2) and Vero-E6 cells were provided by State Key Laboratory of Virology, Wuhan University. Vero-E6 cells were cultured in DMEM (Gibco, Cat.11995–040) supplemented with 10% fetal bovine serum (Gibco, Cat.10091–148) and 1% penicillin and streptomycin solution (BasalMedia, Cat.S110JV) at 37°C with 5% CO2. BHK-21-hACE2…

Continue Reading A potent, broadly protective vaccine against SARS-CoV-2 variants of concern

Nociceptor neurons affect cancer immunosurveillance

Secondary use of biopsies as research specimens The ten melanoma samples used in this study were collected by Sanford Health and classified by a board-certified pathologist. Their secondary use as research specimens (fully de-identified formalin-fixed, paraffin-embedded (FFPE) blocks) was approved under Sanford Health IRB protocol 640 (titled ‘Understanding and improving…

Continue Reading Nociceptor neurons affect cancer immunosurveillance

Cellular glycan modification by B3GAT1 broadly restricts influenza virus infection

Ethics statement All procedures involving laboratory mice were approved by the Duke University IACUC under the protocol numbers A189-18-08 and A142-21-07. Mice were housed with up to five mice per cage and the ambient room conditions ranged from 70–74 °F and 30–70% humidity with a 12-hour dark/light cycle. Animals were assessed…

Continue Reading Cellular glycan modification by B3GAT1 broadly restricts influenza virus infection

Rapid and robust pipeline for developing mRNA vaccines for future SARS-CoV-2 variants

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the causal agent of the ongoing coronavirus disease 2019 (COVID-19) pandemic, has claimed more than 6.5 million lives worldwide. The genomic evolution of the virus has led to the rapid emergence of new SARS-CoV-2 strains, and several of these variants are more infectious and…

Continue Reading Rapid and robust pipeline for developing mRNA vaccines for future SARS-CoV-2 variants

Three-dimensional in vitro model to reproduce COVID-19-induced endothelial dysfunction

In a recent study posted to the bioRxiv* preprint server, researchers developed a three-dimensional (3D) in vitro tissue model to assess the effects of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on the vascular endothelial cells. Study: SARS-CoV-2 spike protein induces endothelial dysfunction in 3D engineered vascular networks. Image Credit:…

Continue Reading Three-dimensional in vitro model to reproduce COVID-19-induced endothelial dysfunction

Inferring and perturbing cell fate regulomes in human brain organoids

Experimental methods Stem cell and organoid culture We used six human iPS cell lines (Hoik1, Wibj2, Kucg2 from the HipSci resource47; 409B2 from the RIKEN BRC cell bank; 01F49i-N-B7 (B7) from Institute of Molecular and Clinical Ophthalmology Basel; and WTC from the Allen Institute) and three human ES cell lines (H1-PAX6YFP…

Continue Reading Inferring and perturbing cell fate regulomes in human brain organoids

Transcription-independent regulation of STING activation and innate immune responses by IRF8 in monocytes

Reagents, antibodies, viruses and cells LMW-Poly(I:C), LPS, 2′3′-cGAMP and DMXAA (InvivoGen); hydroxyurea, camptothecin and mitomycin C (MCE); GM-CSF, Flt3L (peproTech); lipofectamine 2000 (Invitrogen); polybrene (Millipore); RNAiso Plus (Takara); HT-DNA (Sigma); SYBR (BIO-RAD); dual-specific luciferase assay kit (Promega); ELISA kit for murine Ifn-β (PBL); ELISA kits for murine IP-10 (Biolegend) were…

Continue Reading Transcription-independent regulation of STING activation and innate immune responses by IRF8 in monocytes

Characterizing Salmonella Typhimurium-induced Septic Peritonitis in Mice

This protocol describes the induction of Gram-negative monobacterial sepsis in a mouse model system. The model is useful in investigating the inflammatory and lethal host responses during sepsis. Sepsis is a dysregulated host immune response to microbial invasion or tissue damage, leading to organ injury at a site distant from…

Continue Reading Characterizing Salmonella Typhimurium-induced Septic Peritonitis in Mice

Turncoat T cells are at the core of multiple sclerosis and other inflammatory central nervous system disorders

scRNAseq replicates. CD45.1+ P14 cells were adoptively transferred into adult CD45.2+ GP or GFAP:GP mice 24h before i.c. infection with rLCMV-GP33. At least six weeks later, mice were treated with antiCD8a depleting antibody or isotype control. One week after depletion, mice were treated with TAM i.p. for four consecutive days…

Continue Reading Turncoat T cells are at the core of multiple sclerosis and other inflammatory central nervous system disorders

Frontiers | A Mutated Nme1Cas9 Is a Functional Alternative RNase to Both LwaCas13a and RfxCas13d in the Yeast S. cerevisiae

Introduction The clustered regularly interspaced short palindromic repeats (CRISPR)–CRISPR-associated (Cas) protein systems naturally exist in prokaryotes. They are RNA-mediated defense mechanisms to protect bacteria and archaea from invading nucleic acids (Barrangou et al., 2007). CRISPR–Cas systems are classified into two classes (1 and 2). Class 2 CRISPR–Cas require a single…

Continue Reading Frontiers | A Mutated Nme1Cas9 Is a Functional Alternative RNase to Both LwaCas13a and RfxCas13d in the Yeast S. cerevisiae

Postdoc (m/f/d) – CBIRT

Website Leibniz Institute on Aging – Fritz Lipmann Institute (FLI) Job Category: Bioinformatics Job Job ID: 22/16 Date Posted: 05.05.2022 Deadline: 19.06.2022 Location: Jena, Germany The Leibniz Institute on Aging – Fritz Lipmann Institute (FLI) in Jena is a federal and state government-funded research institute and member…

Continue Reading Postdoc (m/f/d) – CBIRT

Figure 3 | Bone Marrow-Derived Cells Contribute to the Maintenance of Thymic Stroma including TECs

Single sorted bone marrow-derived CD45+ EpCAM+ cells express both the CD45 and EpCAM genes and surface proteins. CD45+ EpCAM+ cells were sorted to high purity and analyzed for CD45 and EpCAM expression using immunofluorescence microscopy, as well as qRT-PCR. CD45+ EpCAM- cells were also sorted as a control. (a) FACS…

Continue Reading Figure 3 | Bone Marrow-Derived Cells Contribute to the Maintenance of Thymic Stroma including TECs

RNA interference triggers that target SARS-CoV-2 genome

Coronavirus disease 2019 (COVID-19) vaccines have played a critical role in reducing transmission of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) transmission. However, with emerging reports of waning vaccine efficacy, there remains an urgent need to develop prophylactic measures against COVID-19. In a recent study published on the bioRxiv*…

Continue Reading RNA interference triggers that target SARS-CoV-2 genome

Exploring m6A and m5C Epitranscriptomes upon Viral Infection: an Example with HIV

Hi, in this video we’ll show you how to explore the m6A and m5C epitranscriptomes upon viral infection. To start the experiment, you’ll need 50 million cells for the noninfected and 50 million for the infected condition. In order to obtain a high quality dataset upon analysis, you’ll need to…

Continue Reading Exploring m6A and m5C Epitranscriptomes upon Viral Infection: an Example with HIV

Rat Anti-Mouse CD25/IL-2R Antibody

Name Rat Anti-Mouse CD25/IL-2R Antibody (GWB-31929F) Related Product Names Rat Anti-Mouse CD25/IL-2Ra-PE; PMP-22; Protein CD25; SR13 myelin protein; Schwann cell membrane glycoprotein; SAG Cd25; Pmp-22Rat Anti-Mouse CD25/IL-2RPmp22 Gene Symbol IL2RA Gene Fullname Interleukin-2 receptor subunit alpha Gene id 3559 Swissprot ID P01589 Protein Name Interleukin-2 receptor subunit alpha Protein Accession…

Continue Reading Rat Anti-Mouse CD25/IL-2R Antibody

A cell-based phenotypic library selection and screening approach for the de novo discovery of novel functional chimeric antigen receptors

Cell lines and culture Unless stated otherwise, all tumor target cell lines were sourced from the ATCC via LGC Standards. Human endogenous mesothelin (MSLN)-positive target cell lines H-226 (lung carcinoma, MSLN+, ATCC® CRL-5826™) and AsPC-1 (ATCC CRL-1682), and MSLN-negative Raji control cells (ATCC CCL-86) were cultured in RPMI-1640 Glutamax (Life Technologies,…

Continue Reading A cell-based phenotypic library selection and screening approach for the de novo discovery of novel functional chimeric antigen receptors

Identification of a regulatory pathway inhibiting adipogenesis via RSPO2

Integration of APC scRNA-seq data reveals heterogeneity of adipocyte progenitor cells In a previous study9, we defined Lin−Sca1+CD142+ APCs as adipogenesis regulatory (Areg) cells and demonstrated that these cells are both refractory toward adipogenesis and control adipocyte formation of APCs through paracrine signaling. In contrast, Merrick et. al.4 observed that Lin−CD142+ cells…

Continue Reading Identification of a regulatory pathway inhibiting adipogenesis via RSPO2

The role of ATXR6 expression in modulating genome stability and transposable element repression in Arabidopsis

Significance The plant-specific H3K27me1 methyltransferases ATXR5 and ATXR6 play integral roles connecting epigenetic silencing with genomic stability. However, how H3K27me1 relates to these processes is poorly understood. In this study, we performed a comprehensive transcriptome analysis of tissue- and ploidy-specific expression in a hypomorphic atxr5/6 mutant and revealed that the…

Continue Reading The role of ATXR6 expression in modulating genome stability and transposable element repression in Arabidopsis

Phospholipase A2 inhibitor and LY6/PLAUR domain-containing protein PINLYP regulates type I interferon innate immunity

Significance Interferon (IFN)-mediated antiviral responses serve as the first line of the host innate immune defense against viral infection. Here we identify a previously uncharacterized protein designated phospholipase A2 inhibitor and LY6/PLAUR domain-containing protein (PINLYP), which is essential for embryonic development and plays an important role in type I IFN…

Continue Reading Phospholipase A2 inhibitor and LY6/PLAUR domain-containing protein PINLYP regulates type I interferon innate immunity

IL-2R binders queued for testing

Starting in summer 2021, we ran 6 rounds of IL-2R binder design puzzles. Now we’ve selected 115 promising Foldit designs to test for binding in the lab! In early 2022, we will conduct a FACS experiment to test if these protein designs successfully bind to the IL-2R target. Targeting IL-2R…

Continue Reading IL-2R binders queued for testing

Single-cell delineation of lineage and genetic identity in the mouse brain

STICR lentiviral library preparation and validation We synthesized a high-complexity lentivirus barcode library that encodes approximately 60–70 million distinct oligonucleotide RNA sequences (STICR barcodes). STICR barcodes comprised three distinct oligonucleotide fragments cloned sequentially into a multicloning site within the 3′ UTR of an enhanced green fluorescent protein (eGFP) transgene under…

Continue Reading Single-cell delineation of lineage and genetic identity in the mouse brain

Flow Cytometry and DNA Sequenzing Facility

BD Influx Sorter, a globally unique prototype equipped with fiber optics and octagon/trigon arrays. BD Influx Sorter, a globally unique prototype equipped with fiber optics and octagon/trigon arrays. Flow Cytometry Fluorescence-Activated Cell Sorting (FACS) is a technology that enables qualitative and quantitative analysis of cell mixtures and sorting by individual…

Continue Reading Flow Cytometry and DNA Sequenzing Facility

IRE combined with toripalimab versus IRE alone for LAPC

Introduction Pancreatic ductal adenocarcinoma (PDAC) is a lethal gastrointestinal disease with increasing morbidity, which also has a growing impact on cancer-specific mortality worldwide.1 Nearly 40% of all PDAC cases are localized to the pancreas and characterized with the involvement of major vascular structures, leading to unresectable disease without metastases detected…

Continue Reading IRE combined with toripalimab versus IRE alone for LAPC

MUSC and Helix launch In Our DNA SC, first-of-its-kind population genomics program to drive preventive, precision health care for South Carolinians | MUSC

Large-scale initiative will advance innovative research, improved health outcomes CHARLESTON, S.C. and SAN MATEO, Calif., (Sept. 20, 2021) – The Medical University of South Carolina  (MUSC) and Helix have announced a strategic collaboration to develop a first-of-its-kind population genomics initiative in South Carolina called In Our DNA SC. The large-scale program is designed…

Continue Reading MUSC and Helix launch In Our DNA SC, first-of-its-kind population genomics program to drive preventive, precision health care for South Carolinians | MUSC

Disabling de novo DNA methylation in embryonic stem cells allows an illegitimate fate trajectory

The mammalian genome is characterized by widespread methylation of cytosine residues. After fertilization, however, both maternal and paternal genomes undergo extensive demethylation, reaching a low point in the blastocyst (1⇓⇓–4). The embryo genome is then remethylated by the activity of de novo DNA methylation enzymes (5). Mouse embryonic stem (ES)…

Continue Reading Disabling de novo DNA methylation in embryonic stem cells allows an illegitimate fate trajectory

Serotiny hiring Data Scientist / Bioinformatics Scientist

About Serotiny Serotiny is a company that is engaged in the discovery of proteins for cell therapies. It focuses on such proteins as CARs (chimeric antigen receptors) for the treatment of cancers and genetic disorders. Job Description Description Serotiny invents multi-domain proteins for gene and cell therapies including genome editors…

Continue Reading Serotiny hiring Data Scientist / Bioinformatics Scientist

Defects in 8-oxo-guanine repair pathway cause high frequency of C > A substitutions in neuroblastoma

Significance The collection of large amounts of whole-genome sequencing data allowed for identification of mutational signatures, which are characteristic combinations of substitutions in the context of neighboring bases. The clinical significance of these mutational signatures is still largely unknown. In neuroblastoma, we showed that high levels of cytosine > adenine…

Continue Reading Defects in 8-oxo-guanine repair pathway cause high frequency of C > A substitutions in neuroblastoma

New Results from BabySeq, Pharmacogenomic Updates, New Products

September 1, 2021 | Harvard-Israel and Sema4-Avera launch precision medicine partnerships, Rice announces new Genetic Design and Engineering Center, Foundation Medicine partners with Epic, and Amgen and USC provide cryo-TEMs. Plus new products from BioIVT, BD.   Pluto Biosciences has launched its cloud-based, collaborative life sciences platform that leverages technology…

Continue Reading New Results from BabySeq, Pharmacogenomic Updates, New Products