...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... of Stem Cell Research Plasmids . For more information on stem cells, visit the NIH Stem Cell information... reprogramming of human somatic cells to pluripotency. Cell Stem Cell. 2008 Sep 11. 3(3):346-53. Jaenisch...

... 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...protocol is for suspension cells. Adherent cells can either grow as individual cells in 96-well flat bottom...Deletions in Mammalian Cell Lines CRISPR: Protocol for Genomic Deletions in Mammalian Cell Lines Other Addgene...detail for murine erythroleukemia (MEL) cells, a suspension cell line. The culture medium in all steps ...numerous cell types using preferred culture conditions and transfection strategies for each cell type. While...Ensure there are 2 x 10 6 cells per CRISPR pair. Resuspend 2 x 10 6 cells in 100 μl of electroporation solution...

...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...

...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...116 mEGFP Proliferating cell nuclear antigen (PCNA) DNA replication foci IPSC cell lines can be ordered ...ordered from the Allen Institute for Cell Science Cell Catalog (Link opens in a new window) . Protocols The ...with this plasmid collection. Single cell passaging of human iPS cells (Link opens in a new window) RNP transfection...for Cell Science has deposited plasmids to generate fluorescently tagged proteins of key cellular structures...

...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 ...

...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 ...

...repair protein, and is suitable for both cell-permeant and non-cell-permeant proteins. CLIP-tag CLIP-tag ...Control for cell surface localization 101131 pACP-GPI Control Plasmid ACP/MCP-tag Control for cell surface... sub-cellular localization (membrane, nucleus, etc.) are also available Non-toxic to living cells To learn...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 ...visualization of proteins inside living or fixed cells. The plasmids associated with this technology are...

... 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...

...vectors designed to express nucleases in different cell types. Zinc finger arrays can be transferred in-...vectors designed to express nucleases in different cell types. Zinc finger arrays created using ZFC reagents...nuclease domains: pST1374 (for expression in mammalian cells or zebrafish) pDW1775 (for expression in plants)... Miller et al., Nat Biotech 2007 ) in mammalian cells or zebrafish (can be used as a substitute for pMLM335... Miller et al., Nat Biotech 2007 ) in mammalian cells or zebrafish (can be used as a substitute for pMLM336... Miller et al., Nat Biotech 2007 ) in mammalian cells or zebrafish). pMLM802 (for expression of a ZFN ... Miller et al., Nat Biotech 2007 ) in mammalian cells or zebrafish). pMG10 (a '“2-in-1” ZFN cassette for...

... pH Imaging intracellular pH in live cells (pHRed) Imaging Intracellular pH in Live Cells with a Genetically...SponGee: A Genetic Tool for Subcellular and Cell-Specific cGMP Manipulation. Cell Rep. 2019 Jun 25;27(13):... Tantama ATP QUEEN ATP sensors for live-cell imaging Live cell imaging of metabolic heterogeneity by quantitative...for Imaging Insulin Secretion in Single beta Cells. Cell Chem Biol. 2017 Apr 20;24(4):525-531.e4. Dmytro...accessibility. Cell. 2022 Sep 1;185(18):3390-3407.e18. David Clapham Return to Top Cell Physiology Target...ATP (extracellular) ecAT3.10 extracellular ATP sensor (cell surface targeted) Imaging extracellular ATP...Sensors in Arrayed Islands of Cells for multimodal recordings of cellular physiology. Nat Commun. 2020 ...

... mammalian cells 48880 3xFNLDD/pMXs-I2 iChIP plasmid, expresses 3xFNLDD in mammalian cells 48972 3xFNLDD...mammalian cells 49536 3xFNLDD/pMXs-puro iChIP plasmid, expresses 3xFNLDD in mammalian cells 51240 3xFLAG-dCas9...Thy28-MYH9 axis in B cell-specific expression of the Pax5 gene in chicken B cells. Fujita T, Kitaura F...enChIP plasmid, expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis to purify specific genomic regions...3xFLAG-dCas9/pMXs-puro Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis to purify specific genomic regions...3xFLAG-dCas9/pMXs-IG Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis to purify specific genomic regions...3xFLAG-dCas9/pMXs-I2 Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis to purify specific genomic regions...

... preparations are added to cells in culture. Three to five days later, cell cultures are inspected for... a new vial of early passage cells. Approximately 2 weeks post-thaw, cell culture supernatant is tested...involves transducing cells with serial dilutions of the lentiviral vector, harvesting cells several days later...see maps section for images). Figure 2: AAV Pro cells were transduced with either pAAV-Ef1a-mCherry-IRES-Cre... (55634-AAVrg) alone at 1.7E6 viral genomes (vg)/cell, pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] (Addgene 84445...Production Lentiviral preparations are produced in 293T cells using the 2nd generation psPAX2 and pMD2.G packaging...packaging system developed by the Trono lab . Cell culture medium (OptiPro SFM) containing lentivirus is ...

... and track clones of cells over time. Examples of useful tools for studying cells that share clonal origin...Shendure Lab Simultaneous single-cell profiling of lineages and cell types in the vertebrate brain - Alex...Mumm Lab. NTR 2.0/MTZ-based cell ablation system that allows sustained cell loss and ablation of ...Sauka-Spengler Lab Lineage Tracing Cell lineage tracing in multicellular systems typically relies on genetic...below: UFlip CRISPR targeted integration vectors for cell-type specific conditional gene inactivation - Jeffrey...Pantazis Lab Multibow: digital spectral barcodes for cell tracing - Sean Megason Lab Zebrabow - Albert Pan... previously resistant cell types. CreLite - George Eisenhoffer Lab. An optogenetically controlled...

...hard-to-transfect mammalian cells. In addition, "viral pseudotyping" can be used to target specific cells or tissues. ...genetic material into a cell, and even deliver this genetic material to specific cells in an organism. Figure...Genome Packaging Capacity Virus Size (nm) Cells Infected Target Cell Genome Integration Immune Response Lentivirus...because of their ability to transduce non-dividing cells. These viruses enable long-term stable gene expression...ability to preferentially infect and kill cancer cells. Adenoviruses have high transduction efficiency ... though this infectivity is limited to dividing cells. Retroviruses integrate permanently into the host...for the delivery of this genetic information to cells. Adapting plasmid DNA for use with virus-mediated...

...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 ...information Mouse and Cell Line Repositories (Link opens in a new window) Coriell Institute - cell line repository...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...

... 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...

...find both single-allelic and bi-allelic cells. Single-allelic cells usually make up the majority in culture...allows you to sort for GFP-positive cells and to enrich for those cells that were positively transfected....specificity. A good reference is this Cell paper: http://www.cell.com/abstract/S0092-8674(13)01015-5 As for.... A good example/reference is this Cell paper: http://www.cell.com/abstract/S0092-8674(13)01016-7 In the...CRISPR is used to engineer a locus, isolate single cell colonies, then genotype them, it is possible to ...or delivery the two plasmids together into your cells. If you want to use double nickase, you need to ...each well on 24-well plate, co-deliver into the cells with the Cas9/sgRNAs vector (like PX330). For large...

...effectors to visualize subcellular positioning of target DNA sequences in living cells (TALE-mediated Genome...serial ligation. Validated in zebrafish somatic cells. Keith Joung Zhang Lab TALE Toolbox PCR/Golden Gate...ligation-independent cloning (LIC). Validated in human cells. Veit Hornung Musunuru/Cowan Lab TALEN Kit Kit comprised..., both in microinjected embryos and transfected cells. TALEN expression is driven by the strong CAG promoter...destination vectors and a reporter vector for mammalian cell-based validation assay that are designed for use... the evaluation of TALEN activities in cultured cells. Golden Gate TALEN 2.0 47388 pcDNA3.1-GoldenGate...in mammalian systems including hard-to-transfect cell lines. Features include (i) Esp3I (BsmBI) restriction...

...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 ...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 ...genomic loci in living human cells by an optimized CRISPR/CAS system. Cell . 155 (7), 1479–1491. PMID: ...

...nucleotide variant lines in an isogenic cellular background. All cell lines and phenotypic datasets that result...CRISPR Guide CRISPR Protein Tagging Plasmids for Stem Cell Research Neurodegeneration Research Collection The...Plasmids The following plasmids can be used to create cell lines with endogenously-tagged gene variants or ...or to differentiate human iPSCs into other cell types, as noted in the purpose field: ID Plasmid Description...neurodegenerative diseases with genomic engineering: A new stem cell initiative from the NIH. Neuron . 2021 Apr 7;109...