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We narrowed to 818 results for: cell

Showing: 211 - 220 of 818 results

  1. What's New in CRISPR - March 2020

    Type
    Blog Post
    ...for studying the bacterial cell cycle, asymmetric cell division, and cellular differentiation, but it takes...crescentus Multiplexed gRNA for human pluripotent stem cells CRISPR-based DNA demethylation Optogenetic repressors...Figure 1: Serial dilutions of CRISPRi in Caulobacter cells for ctrA gene expression using various dCas9. Image...lab repressed ctrA, a gene encoding an essential cell cycle regulator. The lab expressed both dCas9 and...piggyBac transposon system for human pluripotent stem cells Lindy Barrett’s lab developed new fluorescently ... of multiplexed gRNAs in human pluripotent stem cells. Previously, CRISPRi or CRISPRa depended on continued...KRAB and dCas9-VPR for quantifying and tracking cells that express both the dCas9 construct and the fluorescently-labeled...

  2. iPSC Neurodegenerative Disease Initiative Plasmid Collection

    Type
    Collection
    ...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...

  3. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

    Type
    Blog Post
    ... engineer cells for specific purposes. Unfortunately, plasmid copy numbers vary within cell populations...problems. Rapidly dividing cells initiate genome replication more than once per cell division and thus, genes.... When cells are dividing rapidly, multiple rounds of replication are initiated before the cell has divided...growth rate can all impact plasmid copy number in a cell. One can attempt to eliminate fluctuations due ... pSC101 with copy numbers ranging from 3-100 per cell, a stabilized promoter buffered against the effects... to stationary phase, the number of plasmids per cell increases 4-5 fold (for pUC plasmids) or ~2 fold...fold (for p15a, R6K, and ColE2 plasmids) as cell division slows (Kittleson et al., 2011). Segall-Shapiro...

  4. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    ...illumination of neighboring cells evokes action potentials (APs) in patched cells expressing wild type CoChR...techniques for live-cell imaging of the dynamic changes in biochemical activity within the cell. Their Fluorescence...28646204. soCoChR: A novel opsin that enables single-cell optogenetics Article contributed by Meghan Rego ...wild type CoChR, soCoChR localizes primarily to the cell body with very little signal detected along the ...CoChR, these APs are absent in soCoChR-expressing cells. Finally, using synaptic receptor blockers to distinguish...precise mapping of neuronal networks at the single-cell level. Various soCoChR plasmids described in this...strain to assay for functional decaffeination in cells. The ΔguaB E. coli are missing an essential enzyme...

  5. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    ... expressed in cells via plasmids or by the creation of stable cell lines and these cells are then grown... complex. Not performed in a native cell context. For AP-MS, cells are lysed before the bait protein is...and interacting proteins in mammalian cells. The Journal of cell biology. PubMed PMID: 22412018. PubMed...against PPIs that aren't stable outside the native cell context. The Split-BioID method Enter Split-BioID...tagging of proteins in both BioID or Split-BioID, cells are lysed, protein extracted, and labeled protein...proteins that interact with the bait in a native cell contact. Other PPI methods may require protein interactions...interactions to remain stable (or to be stabilized) during cell lysis and lysate fractionation. Important considerations...

  6. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    ... do so, their T-cells were grown in vitro, their mutations corrected, and the T-cells were transferred...functioning T cell repertoire (1). In this example, it was possible to transduce the T-cells outside the...a wide range of cell types including but not limited to muscle, liver, and brain cells. There are a multitude...chain (c) and is characterized by a lack of immune cell development and function. To date, gene therapy ...that can deliver gene therapies directly to the cells that need them. Adeno-associated virus (AAV) – based...delivery of nucleic acid cargo (DNA or RNA) to target cells has been challenging. A variety of vector delivery...expression over many years in postmitotic, long-lived cell types. AAVs in the clinic Thus far, AAV vectors...

  7. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    ...large amounts of carbohydrates that can inhibit the cell lysis process. To ensure that lysis is complete,...strains adapted for protein purification use the cell’s resources to produce high levels of protein, not...difference? In the simplest of terms, more volume = more cells = more plasmid. Plasmids can vary in copy number...of plasmid copies supported within each bacterial cell in your culture. Copy number is partially specified...richer media or adding supplements can improve your cell density, leading to an improved plasmid DNA yield... supplements to the media You can achieve higher cell density using a nutrient-rich broth such as Terrific...glucose (in moderation) may also serve to increase cell density and plasmid yield. Start with a single, ...

  8. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    ...same: Deliver CRISPR library to cells of interest. Select for cells of interest (positive or negative...infect specific cell or tissue types. Using serotypes that have a known tropism for your cells of interest... a host cell’s genome. In this state, AAV cargo can be stably expressed in non-dividing cells for long...mice are involved. Ex vivo starts with transducing cells with the CRISPR library in vitro before transplanting...interest — for example, AAV8 for kidney cells — can help improve infection rates in your experiment. ...AAV vectors, enhancing their affinity for specific cell and tissue types. The combination of serotypes and...integration sites seem to only occur in the target cells or tissues. AAV also shows improved transduction...

  9. Viral Production at Addgene

    Type
    Blog Post
    ... 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 two weeks post-thaw, cell culture supernatant is tested...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...preparations ("-LV” catalog items) 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 first...modified from Wang et al., 2018). Briefly, 293T cells are transduced with serial dilutions of a lentiviral...

  10. CRISPR Plasmids - Tagging

    Type
    Collection
    ...Drosophila cells. N terminal tagging in Drosophila cells 3.2 MB C terminal tagging in Drosophila cells 3.3 MB...following article: Natsume, et al. Cell Reports 2016 Förstemann Drosophila Cell Tagging System The Förstemann... Allen Institute for Cell Science Plasmid Collection The Allen Institute for Cell Science has produced...tagged, human, induced pluripotent stem cell (hiPSC) lines. These cell lines were created using CRISPR and...tagging transcription factors with FLAG in human cell lines. To repeat the tagging in your system, use... from the Doyon Lab’s publication: Dalvai et al. Cell Rep. 2015 . Alternatively, cDNAs can be cloned directly... CRISPR tagging technique for use in Drosophila cells that uses PCR to generate both an expression cassette...
Showing: 211 - 220 of 818 results