We narrowed to 819 results for: cell
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TypeGuide...-activated cell sorting (FACS) is used to sort cells expressing GFP and later, sorted cells are expanded...actively dividing cell types, whereas gamma-retroviruses can only infect mitotically active cell types. This...Using Packaging Cell Lines Gamma-retroviral vectors can be packaged using helper-free cell lines (Figure...-retroviral packaging cell line developed by Garry Nolan at Stanford. These cell lines contain gag-pol...and rat cells), or an amphotropic envelope, Phoenix-AMPHO (for the infection of mammalian cells). Using...make stable cell lines, VSV-G is often put under an inducible promoter. Depending on the cell type, there... — T and B cell transduction Murine Leukemia Virus (MuLV) and HIV glycoprotein — CD4+ cell transduction...
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Plan Your Experiment
TypeGuide...difficult-to-transfect cells. You can direct expression to specific cell types by using cell-specific promoters...human embryonic kidney 293 (HEK293) cells. For more difficult cell types, you may need to look into other...are good choices for more difficult cell types like primary cells. For CRISPR delivery, lentiviral and...deliver to target cells through microinjection or electroporation. The target cells translate the mRNA...complexes directly into your target cells. In this case, your target cells are not transcribing DNA to mRNA...possible genotypes within the resulting cell population. Some cells may remain wild type due to either (1...modifications . Cell Population The first step of validation is determining the type of cell population you... -
Lentiviral Vector Guide
TypeGuide...production cell line, typically human embryonic kidney 293T (HEK293T) cells. Standard HEK293T cells are sufficient...fluorescence-activated cell sorting (FACS) is used to sort cells expressing GFP and later expanded into a cell line. More...in a variety of cell types and models. Lentiviral vectors can target non-dividing cells, enable stable ...actively dividing cell types, whereas gamma-retroviruses can only infect mitotically active cell types. This...transfected into HEK293T cells using your preferred transfection method. The HEK293T cells will produce the viral...make stable cell lines, VSV-G is often put under an inducible promoter. Depending on the cell type, there...clinical tools. Stable Cell Lines Lentiviral vectors can be used to make stable cell lines due to their integration... -
Adeno-associated virus (AAV) Guide
TypeGuide...based on other human cell lines, such as HeLa and A549 cells, and even insect Sf9 cells , have also been ...able to infect cells and replicate autonomously. Human embryonic kidney 293T (HEK293T) cells are the most...HEK293T cells are then extracted and purified for their use as viral vectors to deliver genes to cells in ...persist in non-dividing cells for a period of years without damaging the host cell. Episomal stability allows...broad range of cell types in vivo. AAV-DJ has been optimized further for specific cell and tissue types...co-delivered to the host cells. The full-length transgene is later reconstructed within the cell via different mechanisms...main cell body, or soma, towards the synapse at the axons. Blood-brain barrier (BBB) Layer of cells that... -
Antibody Guide
TypeGuide... produced. Cells - Antibodies can also be produced in cell cultures using hybridomas, cell cultures created...created by fusing B cells with myeloma cells (immortalized B cells). The genetic reorganization that creates...identify and sort cells from a mixed population. Overview of application Collect cells from cell culture or ...specificity occurs in the B cell itself prior to antibody production; therefore, each B cell only produces one ... specific hybridoma cells into plasmids. Plasmids are then transfected into cells and induced to express...determines the isotype of the antibody. B cells, the immune cell responsible for producing antibodies, can...mature B cell can clone itself to enable higher production of a single antibody, but each B cell can only... -
Arf GTPase Family
TypeCollection...concepts and challenges. Molecular Biology of the Cell , 30 (11), 1249–1271. PMID: 31084567 (Link opens... -
TALEN Guide
TypeCollection.... Targeted gene disruption in somatic zebrafish cells using engineered TALENs. Sander JD, Cade L, Khayter... -
Science Careers: Unruly Interests Feed Many Paths
TypeBlog Post...sciences, viewed through the lens of development, cell, and molecular biology. Beyond that, she is also... -
FPbase: A new community-editable fluorescent protein database
TypeBlog Post...Associate at Harvard Medical School. He manages the Cell Biology Microscopy Facility, specializing in advanced... There are many online guides – including an excellent series of posts by Joachim Goedhart on the Addgene... -
In Vivo Biotinylation of Bacterial Fusion Proteins
TypeBlog Post... Eye Institute and is particularly interested in cell signaling. Additional Resources on the Addgene...successfully used this plasmid to biotinylate the extracellular domain of the erythropoietin receptor and subsequently... -
New and Upcoming Viral Vectors - September 2019
TypeBlog Post...recommend validating the serotype in your target cells before conducting the experiment. Likewise, a lot... -
Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation
TypeBlog Post... small amount of spurious DNA from the packaging cell line, helper and packaging plasmids. We decided ... -
Multifunctional Neural Fibers: Modulation from Gut to Brain
TypeBlog Post...and chemically excited a class of enteroendocrine cells within the intestines known as neuropods. They observed... -
Three Tips for Preventing Viral Plasmid Recombination in Your Samples
TypeBlog Post...efficient genome editing in Escherichia coli. Microb Cell Fact 19. https://doi.org/10.1186/s12934-020-01321... -
Unique and persistent IDs for improved reproducibility: Addgene now supports RRIDs and compact identifiers for all plasmids
TypeBlog Post...example, several journals like eLife, Nature, and Cell use Research Resource identifiers (RRIDs) which... -
Identifying Sequence Elements with SnapGene's Feature Database
TypeBlog Post..., and also a Professor of Molecular Genetics and Cell Biology at the University of Chicago. He was an ... -
Plasmids 101: TOPO Cloning
TypeBlog Post... Transform TOPO Cloning Reaction into Competent Cells: You can use your standard lab protocol for this... -
Even more elegant: Single injection CRISPR/Cas9 in C. elegans
TypeBlog Post...systems may prove effective in other species or in cell culture. In addition to CRISPR/Cas9 applications... -
History of CRISPR Cas - A tale of survival and evolution
TypeBlog Post...devise new strategies to reproduce within a host cell. A few virus particles have learned to circumvent... -
Guide to Using Pooled Libraries
TypeGuide...200x the number of cells as plasmids in the TKO library. Using large numbers of cells minimizes the effects...selection Most cells die or do not pass selection (in the case of a reporter) Sort “winning” cells Sequence ...library Apply selection; most cells will survive Perform NGS on the surviving cells Compare NGS results between...identify those cells that do not survive the selection mechanism. You’ll infect two sets of cells and subject...libraries, plasmid DNA can be delivered directly to the cells of interest. With others, notably pooled lentiviral...subsequently used to deliver the plasmids to the cells of interest. In either case, next-generation sequencing...Pooled Library Screens In pooled library screens, cells are infected at a very low multiplicity of infection...