...Pre-implantation Blastocyst Cell Types from Fibroblasts. Cell Stem Cell. 2019 Jun 6. Buganim Glial Cells Retinal Ganglion... Plasmids Stem Cell Research Plasmids Stem cells are proliferative, unspecialized cells with the capacity...different cell types in the body during development. Unlike other cell types, each stem cell has the potential...generate different cell lineages. Pluripotent stem cells, such as embryonic cells, can potentially give...any other cell type in the body. Multipotent stem cells, such as adult (or somatic) stem cells, can differentiate...disease-specific cells. For this reason, iPSCs have become an excellent unlimited stem cell source for therapeutic... of Stem Cell Research Plasmids . For more information on stem cells, visit the NIH Stem Cell information...

... Depositor Collections Cell Migration Consortium (CMC) Plasmids Cell Migration Consortium (CMC) Plasmids...collaboration between the Cell Migration Consortium and Addgene. Member laboratories of the Cell Migration Consortium...This resource is a collaboration between the Cell Migration Consortium (CMC) and Addgene, which highlights...Consortium have deposited plasmids useful for cell migration research at Addgene. CMC investigators who have...MIT Rick Horwitz , University of Virginia Denise Montell , University of California, Santa Barbara Martin...

...obtain approximately 30 cells per 96-well plate. Include 100 μl per well of cell culture media. For the...is for suspension cells. Adherent cells can either grow as individual cells in 96-well flat bottom plate... While MEL cells are suspension cells, instructions for adherent cells have also been included. Ensure... Fluorescence Activated Cell Sorting (FACS) of Transfected Cells Prepare cells for FACS by filtering them...limiting dilution at 30 cells per 96-well round-bottom plate. Optimize plating the cell type used at limiting...parental and bulk sorted cells by resuspending parental and bulk cell pellets in 50 μl of DNA extraction...suspension cells, transfer all clones to a single 96-well plate that already contains 50 μl cell culture ...

...Consortium plasmids that encode the extracellular regions of cell surface receptor and secreted proteins...expression plasmids that encode the extracellular regions of cell surface receptor and secreted proteins...screens or cellular assays to identify novel interactions or pathways that are important in cellular recognition...AVEXIS (for Avidity-based extracellular interaction screen) in which the extracellular regions of receptor ...and Wright GJ Molecular & cellular proteomics 2010 Dec; 9(12):2654-65. A cell surface interaction network...screening for novel low-affinity extracellular protein interactions. Bushell KM, Söllner C, Schuster-Boeckler... Plasmid Collections EXPRESs Plasmids EXtracellular Plasmid RESource (EXPRESs) Consortium You may...

...give the cell its shape, organize cellular organelles, and are necessary for movement and cell division...these well-characterized proteins. General Structures The cell contains a variety of organelles and subcellular...between animal cells. They are important for holding cell structures together and reducing cell stress. There...Collections Fluorescent Proteins Organelle Markers Fluorescent Proteins: Subcellular Localization More Fluorescent...proteins that are targeted to various organelles or subcellular structures. You should fuse your protein...example, houses the cell’s genetic material. The mitochondria produces the energy cells required for chemical...the table below to find markers of general cellular organelles and structures. ID Plasmid Structure Targeting...

...for Cell Science has deposited plasmids to generate fluorescently tagged proteins of key cellular structures... Protein Guide Subcellular Localization Plasmids Protocols The Allen Institute for Cell Science has produced...fluorescently tagged proteins of key cellular structures in human cell lines of their choice. For a full ...Tagging in Human Cells on the Addgene Blog. Plasmids for Endogenous Gene Tagging in Human Cells Find additional...additional cell line information from the Allen Institute for Cell Science data portal (Link opens in a new...images from the hiPS cell lines gRNA sequences used in the creation of the hiPS cell lines The following...plasmids can be used to create cell lines with endogenously-tagged cellular structures. Addgene ID Plasmid...

... sub-cellular localization (membrane, nucleus, etc.) are also available Non-toxic to living cells To learn...repair protein, and is suitable for both cell-permeant and non-cell-permeant proteins. CLIP-tag CLIP-tag ... (CoA). These tags are particularly well suited for labeling cell-surface proteins, and are useful for...Control for cell surface localization 101131 pACP-GPI Control Plasmid ACP/MCP-tag Control for cell surface...visualization of proteins inside cells. Depositor... Depositor Collections New England Biolabs Cell-Imaging Plasmid Collection...Collection New England Biolabs Cell-Imaging Plasmid Collection Technology Plasmids Supplemental Information ...

...foreign antigens to adaptive immune cells, such as T cells and B cells. About a week following the initial...presenting cells. B cells in secondary lymphoid organs recognize specific antigens through the B cell receptor...have been described. NK Cell Signaling Natural killer (NK) cells can kill cells that have been infected... within an infected cell membrane, triggering cell death. TCR Signaling The T cell receptor (TCR) engages...through effects on numerous immune cell types including T cells and B cells. It also impacts the development...function of innate immune cells including NK cells, macrophages, dendritic cells, and granulocytes. TLR ...

...ATP (extracellular) ecAT3.10 extracellular ATP sensor (cell surface targeted) Imaging extracellular ATP... pH Imaging intracellular pH in live cells (pHRed) Imaging Intracellular pH in Live Cells with a Genetically...indicators FR-GECO1 Robert Campbell Lab (unpublished) Robert Campbell Calcium Yellow protein calcium sensor...SponGee: A Genetic Tool for Subcellular and Cell-Specific cGMP Manipulation. Cell Rep. 2019 Jun 25;27(13):...designed to specifically localize to cellular compartments or organelles. Addgene’s database includes many... vitro vs. in vivo , cytoplasmic vs. extracellular vs. organelle-localized). Browse or search the tables...STIM1 signalling system regulates directed cell migration. Nat Cell Biol. 2014 Feb;16(2):133-44. Tobias Meyer...

... enhancer element Gordon Fishell 83894-AAV2 pAAV-hDlx-Flex-dTomato-Fishell_7 Cre recombinase-dependent... enhancer element Gordon Fishell 83894-AAV5 pAAV-hDlx-Flex-dTomato-Fishell_7 Cre recombinase-dependent... enhancer element Gordon Fishell 83894-AAV9 pAAV-hDlx-Flex-dTomato-Fishell_7 Cre recombinase-dependent... enhancer element Gordon Fishell 83894-AAVrg pAAV-hDlx-Flex-dTomato-Fishell_7 Cre recombinase-dependent...hDlx enhancer element Gordon Fishell 83895-AAV1 pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP...hDlx enhancer element Gordon Fishell 83895-AAV2 pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP...hDlx enhancer element Gordon Fishell 83895-AAV8 pAAV-hDlx-Flex-GFP-Fishell_6 Cre recombinase-dependent GFP...

...with Haploid Embryonic Stem Cells Carrying a Guide RNA Library. Cell Stem Cell . 17 (2), 221–232. PMID: ...in the same cell. This is called multiplexing , and increases the likelihood that any cell containing ... the cell along with the gRNA(s) and Cas enzyme. The donor DNA contains the desired edit as well as additional...can increase efficiency. Synchronizing cells or modulating the cell cycle can help as HDR takes place during...expressing stable cell line. After applying your screening conditions to your transduced cell population, you...tracking of RNA in live cells, including orthogonal RNA/DNA labeling in the same cell. Figure 11: Overview...promiscuity is unique to bacterial cells and does not occur in mammalian cells. SHERLOCK , developed by the ...

... 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...

...expressed in your target cells. For easy-to-transfect cell types (e.g. HEK293 cells), transfection with standard... CRISPR machinery. For more difficult cells (e.g. primary cells), viral delivery of CRISPR reagents may...identify and enrich positive cells, or selection marker to generate stable cell lines (for details, see Plasmids...wide variety of mammalian cell lines Useful for difficult to transfect cell types, can also be used in...possible genotypes within the resulting cell population. Some cells may be wild type due to either (1) a ...efficient target cleavage in cells expressing both Cas9 and gRNA. Edited cells may be homozygous or heterozygous...will use the example of CRISPR/Cas9 in mammalian cells, many of these principles apply to using CRISPR ...

...spread from this cell to synaptically connected cells (meaning, it can spread to cells that are projecting...infect a single "starter" cell, but new virus cannot spread to any neighboring cells. To enable monosynaptic...provided to the starter cell in trans , via delivery by an AAV. Since this starter cell now has all the viral...projecting to the starter cell). Once in the secondarily infected cells that lack the G protein, the virus ...connected cells. To further refine the targeting of G-deleted rabies to specific starter cells, the tropism...G-protein is delivered to the starter cell. (2) The starter cell now expresses TVA and G-protein. EnvA-pseudotyped...is delivered to the starter cell, and specifically targets the starter cell via the EnvA receptor. RABV...

...for one well in 24-well plate. A good example/reference is this Cell paper: http://www.cell.com/abstract... 10uM oligo stock for each well on 24-well plate, co-deliver into the cells with the Cas9/sgRNAs vector... vector, for a one well in a 24-well plate. A good reference is this paper in Cell . Click here for a ...find both single-allelic and bi-allelic cells. Single-allelic cells usually make up the majority in culture...specificity. A good reference is this Cell paper: http://www.cell.com/abstract/S0092-8674(13)01015-5 As for...allows you to sort for GFP-positive cells and to enrich for those cells that were positively transfected....CRISPR is used to engineer a locus, isolate single cell colonies, then genotype them, it is possible to ...

...channelrhodopsin-2 driven by PV+ basket cell-targeting enhancer Gordon Fishell Viviana Gradinaru 213940-PHPeB ...expressing dTomato driven by PV+ basket cell-targeting enhancer Gordon Fishell Viviana Gradinaru 213937-PHPeB ...channelrhodopsin-2 driven by chandelier cell-targeting enhancer Gordon Fishell Viviana Gradinaru 213936-PHPeB ...expressing dTomato driven by chandelier cell-targeting enhancer Gordon Fishell Viviana Gradinaru 213915-PHPeB ...Collection The Armamentarium for Precision Brain Cell Access is developing a collection of molecular genetic... reagents to gain access to many different brain cell types. The expanded collection of Armamentarium ...10aa-H2B-WPRE3-BGHpA (Alias: CN4496) Expression of SYFP2 (yellow fluorescent protein) plus separate nuclear FLAG...

...Control Boyden 83900 pAAV-mDlx-GFP-Fishell-1 Dlx GFP Control Fishell 99130 pAAV-mDlx-NLS-mRuby2 Dlx mRuby2...Control Fishell 213829 pAAV_BiCHATe27_dTomato_nlsdTomato BiCHATe27 dTomato/nlsdTomato Control Fishell 213855...Control Fishell 213936 pAAV_BiPVe4_dTomato_nlsdTomato BiPVe4 dTomato/nlsdTomato Control Fishell 213940 ...Control Fishell 213944 pAAV_BiSSTe4_dTomato_nlsdTomato BiSSTe4 dTomato/nlsdTomato Control Fishell Chemogenetics...Activator ChR2 Fishell 213830 pAAV_BiCHATe27_ChR2_mCherry BiCHATe27 Activator ChR2 Fishell 213856 pAAV_BiVIPe4...Activator ChR2 Fishell 213915 pAAV_BiLAMP5e3_ChR2_mCherry BiLAMP5e3 Activator ChR2 Fishell 213937 pAAV_BiPVe4... Activator ChR2 Fishell 213941 pAAV_BiPVe3_ChR2_mCherry BiPVe3 Activator ChR2 Fishell 213945 pAAV_BiSSTe4...

...PRKCG MTH Spinocerebellar ataxia 30 Frederic Mushinski 8418 pLTR PKC gamma PRKCG Spinocerebellar ataxia ...GFP CMV Spinocerebellar ataxia 27 Tobias Meyer 21204 GFP-N2-PKCgamma PRKCG GFP CMV Spinocerebellar ataxia...GFP CMV Spinocerebellar ataxia 25 Tobias Meyer 21215 GFP-C2gamma PRKCG GFP CMV Spinocerebellar ataxia ...CMV Spinocerebellar ataxia 23 Bernard Weinstein 21237 PKC gamma dNPS PRKCG HA CMV Spinocerebellar ataxia... CMV Spinocerebellar ataxia 21 Bernard Weinstein 21239 PKC gamma DN PRKCG HA CMV Spinocerebellar ataxia...OPTN His CMV ALS Jon Ashwell 23051 pDEST26-hOPTN-D474N OPTN His CMV ALS Jon Ashwell 23052 pDEST15-hOPTN...hOPTN OPTN GST T7 ALS Jon Ashwell 23053 pDEST17-hOPTN OPTN His T7 ALS Jon Ashwell 23054 pDEST15-hOPTN-D474N...

...genomic loci in living human cells by an optimized CRISPR/Cas system. Cell . 155(7):1479-91. PMID: 24360272... for Mapping Multiprotein Complexes in Human Cells. Cell Rep . 13(3):621-33. PMID: 26456817 Fu Y, Sander...Sontheimer EJ, Maxwell KL, Davidson AR. 2016. Naturally occurring off-switches for CRISPR-Cas9. Cell . 167(7)...described CRISPR-based genome editing in mammalian cell culture. Five years later, PubMed lists more than...orthogonality, and multiplexibility in various species, as well as the development of new applications. The first...multiple loci can be easily expressed in the same cell or organism. The ability to precisely target CRISPR...Current libraries are available for gene knockout, as well as transcriptional activation or repression. Combined...

...information Mouse and Cell Line Repositories (Link opens in a new window) Coriell Institute - cell line repository...Unpublished Joost Gribnau Human Cell Line Models Induced pluripotent stem cells (iPSCs) are a powerful system...opens in a new window) LUHMES cell line , an immortalized pre-neuronal cell line that can be differentiated... RSRT cell line collection (186.8 KB) . To order, please email [email protected] The following cell lines ...Collection and Resource Center MECP2 Animal Models Human Cell Lines CRISPR Tools Plasmids Resources The (Link ...X inactivation results in approximately half of cells expressing wild-type MECP2 and half expressing mutant...treat Rett syndrome and restore functional MECP2 in cells. Search (Link opens in a new window) JAX Mice for...