...suitable for virus purification and isolation of cells, organelles, lipoproteins, and macromolecules. Importantly..., iodixanol is inert and non-toxic to mammalian cells. Therefore, and unlike other density gradient media...

...of our protocols supports reproducibility and accelerates science. Here, we list the specific equipment...shaker ELISA plate washer (optional) Scale Microsoft Excel or similar software Reagents Purified antigen Purified...

...restriction digests needed in molecular biology, and thaw cell lines. There are many different types of water baths...

...min at 12,000 g. Notes: Pellet contains proteins, cell fragments, salt and other extra particles from solutions...

...Note: (Link opens in a new window) Primer3 is an excellent resource for choosing primers. Place thin-walled...

...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...SponGee: A Genetic Tool for Subcellular and Cell-Specific cGMP Manipulation. Cell Rep. 2019 Jun 25;27(13):...STIM1 signalling system regulates directed cell migration. Nat Cell Biol. 2014 Feb;16(2):133-44. Tobias Meyer...Sensors in Arrayed Islands of Cells for multimodal recordings of cellular physiology. Nat Commun. 2020 ...imaging in live cells (CALWY) Genetically encoded FRET sensors to monitor intracellular Zn(2+) homeostasis... Tantama ATP QUEEN ATP sensors for live-cell imaging Live cell imaging of metabolic heterogeneity by quantitative...

... 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...

...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 ...editing systems by manipulating cellular determinants of editing outcomes. Cell . 184 (22), 5635-5652.e29. ...

...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...

...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.... A good example/reference is this Cell paper: http://www.cell.com/abstract/S0092-8674(13)01016-7 In the...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 ...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...

...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...the MECP2 protein in mouse and rat models have accelerated our understanding of Rett syndrome at the molecular...

... 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 ...

...carrying version 1 of the CellTag barcoding library. Version 1 of the Celltag library contains 19973 barcodes...barcodes to combinatorially index cells for single-cell analysis of clonal dynamics. This library expresses...carrying version 2 of the CellTag barcoding library. Version 2 of the Celltag library contains 4934 barcodes...carrying version 3 of the CellTag barcoding library. Version 3 of the Celltag library contains 5737 barcodes...Libraries ID Name Description PI 115643 Morris CellTag Barcode Library Version 1 Ready-to-use lentiviral...

...Sauka-Spengler Lab Lineage Tracing Cell lineage tracing in multicellular systems typically relies on genetic... 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 ...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...

...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...described CRISPR-based genome editing in mammalian cell culture. Five years later, PubMed lists more than...multiple loci can be easily expressed in the same cell or organism. The ability to precisely target CRISPR...genome edits or double-strand breaks. In mammalian cells, Cas13-ADAR2 does not require a specific flanking...proteins can be used to image genomic loci in live cells. RNA targeting : Cas13 enzymes target RNA rather...non-specific cleavage does not occur in mammalian cells. Purify : Epitope-tagged dCas9 can also be used ...

...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...

...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...