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We narrowed to 813 results for: cell

Showing: 221 - 240 of 813 results

  1. Using CRISPR/Cas9 to Edit Disease Out of the Genome

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    Blog Post
    ...Cas9 in Intestinal Stem Cell Organoids of Cystic Fibrosis Patients." Cell Stem Cell. 13, 653-658 (5 December...CRISPR in adult stem cells has huge potential, because the genomes of adult stem cells in organoid cultures...Genetic Disease in Mouse Via Use of CRISPR/Cas9." Cell Stem Cell. 13, 659-662 (5 December 2013). 8. Gerald Schwank...homologous recombination in cultured intestinal stem cells of patients with cystic fibrosis (8).   The difference...applied CRISPR to disease correction in adult stem cells isolated from two patients with cystic fibrosis....is a big advantage over induced pluripotent stem cells, which need to go through a phase of crisis where...B. 2021. “CRISPR/Cas gene therapy.” Journal of cellular physiology.  https://doi.org/10.1002/jcp.30064...

  2. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    ...Virus (MoMLV, MMLV, MuLV, or MLV) or Murine Stem Cell Virus (MSCV) genomes whereas lentiviral vectors ...things) gene delivery, generating pluripotent stem cells, and developing gene therapies for monogenic diseases...vectors, their infectivity is limited to dividing cells. Find tips on using gamma-retroviral vectors in ...are able to transduce dividing and non-dividing cells. More detailed descriptions of the lentiviral elements... to express siRNAs of their choice in mammalian cells.  Learn more about Mammalian shRNA Tools for RNAi...-optimized Cas9 under the control of Murine Stem Cell retrovirus promoter for mammalian gene knockdown...96 retroviral plasmids (based on the Murine Stem Cell Virus, MSCV) each with a unique 6 nucleotide barcode...

  3. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    ...vectors (right) are transduced to target cells. Within target cells, Cas9 machinery is transiently expressed...in order to facilitate its entry into the target cells (Beatty et al., 2012). These features make AdV an... complexes into transformed and non-transformed cells yielded rates of targeted mutagenesis similar to...nuclease-induced gene disruption frequencies in the various cell types ranged from 18% to 65% (Maggio et al., 2014... leads to a vast majority of AdV-modified human cells being subjected to scarless homology-directed genome...therapeutically relevant, non-transformed mammalian cells. These viral vector systems also work equally well... dividing and quiescent, post-mitotic mammalian cells. Applications of CRISPR delivery by adenoviral ...

  4. What's New in CRISPR - March 2020

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    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...

  5. Fluorescent Proteins 101: Luciferase

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    Blog Post
    ...luciferase assays require cell lysis as the most efficient means to disrupt the cell membrane and deliver ...miRNAs for effectiveness. Analyze Cell Signaling Pathways Known cellular signaling pathways can be investigated...luciferase as a genetic reporter in plant and mammalian cells. Luciferase assays have since become a gold standard... serves as an indicator of transcription within cells, where detection of the reporter protein or its ... into the desired mammalian, plant or bacterial cells. Expression of luciferase as the reporter gene is...can permit measurements of luminescence from live cells. Monitor miRNA knockdown Expression of luciferase...codon, which is then transfected into the desired cells. A hybrid mRNA transcript containing luciferase ...

  6. 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...

  7. 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...

  8. 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...

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

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    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...

  10. 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, ...

  11. Lentiviral Prep Service

    Type
    Collection
    ...barcodes to combinatorially index cells for single-cell analysis of clonal dynamics. This library expresses...carrying version 1 of the CellTag barcoding library. Version 1 of the Celltag library contains 19973 barcodes...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...

  12. 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...

  13. 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...

  14. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ...number of cells have been edited. For many experiments, the end goal is to generate a clonal cell line, so...designed your gRNA and introduced it into your target cells with Cas9. Hooray! Now it’s time to make sure your...blog post we’ll explain how to verify that your cells were appropriately edited for your desired mutation...important to know a few things about your target cells and the desired edits. First, you will need to know.... Validating knockout mutations When a knockout cell line is desired, any mutation that introduces a ...input is sequencing trace files of WT and edited cell lines. Then decomposition analysis is performed ...both an unedited population and your Cas9-targeted cells. Simply upload both trace files to the TIDE tool...

  15. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    ...scFvs and T cell receptors and are expressed on T cells to make CAR T cells. The extracellular scFv portion...Bispecific T-cell engagers (BiTE®s), bind CD3 on T cells and a tumor-specific antigen, it brings T cells to a ...scFv part of the receptor can activate T cells to kill cancer cells. scFv’s small size also allows for their...recognizes an antigen of interest while the intracellular T cell receptor portion of the CAR facilitates ... display libraries screen antibody genes from B cells of immunized animals, but it's possible to avoid...also allows for screening scFv generated from the B cell antibody genes from diseased or vaccinated humans...produce scFvs in mammalian, yeast, plant, and insect cells, they are most often expressed from a plasmid in...

  16. Your Top Requested Plasmid in 2016!

    Type
    Blog Post
    ...both SpCas9 and your gRNA in a variety of mammalian cell types. Purpose  SpCas9 This plasmid expresses Streptococcus...the Zhang lab has tested in a variety of mammalian cell lines. Cas9, of course, is the key component of ... puromycin resistance gene enables selection of cells with the plasmid post transfection. The 2A "self...can be used to transfect a variety of mammalian cells and can be modified for viral trasduction (see the...resistance gene that made it less effective in certain cell lines. This point mutation has been corrected in... your gRNA of interest in a variety of mammalian cell lines. Alternative forms of this vector contain ...Common Ways to Deliver CRISPR Plasmids to Mammalian Cells Resources on the Addgene Website Visit our CRISPR...

  17. Important Considerations When Using AAVs

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    Blog Post
    ...transducing the cell type under study. Many AAVs will show significant expression in a variety of cell types, ...optogenetics as tools to investigate the roles of certain cell types in locomotion. We have relied heavily upon...deliver chemogenetic/optogenetic constructs into our cells of interest. Although they have a small packaging...of serotype (which determines which tissues and cells an AAV can infect) can influence an AAV’s value ... tissue? Location (2): Where in the tissue/which cell type? Location (3): Traveling from the original ...AAV5, AAV6, AAV9 Pancreas AAV8 Photoreceptor Cells AAV2, AAV5, AAV8 RPE (Retinal Pigment Epithelium...-vectors/aav/aav-guide. Where in the tissue/what cell type? In the CNS, for instance, there are marked...

  18. Empty Backbones - Choosing Your Perfect Plasmid Backbone

    Type
    Collection
    ...and display of proteins on the extracellular surface of S. cerevisiae cells His-tagged versions of pFastBac... the elements needed for replication in the host cell and are designed for a user to insert a gene of ..., or express the insert they carry in the target cell. These vectors allow you to test the function of....1-mCherry - Expresses mCherry tag in mammalian cells pBI-MCS-EGFP - Tet-inducible Find more Tet-inducible...experiments, scientists often want to create stable cell lines in which the expression cassette of interest...is incorporated into the host DNA. For mammalian cells, you can do this through viral vector delivery...transduce both dividing and nondividing cells, integrate into host genome pLenti CMV GFP DEST ...

  19. Optogenetics Plasmid Collection

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    Collection
    ... optical switch tools to measure and manipulate cells and their biomolecular processes....to blue light and allows cations to flow into the cell. Variants of channelrhodopsin have been engineered...improve the expression and membrane targeting in host cells. Other microbial opsins, such as the Halorhodopsins...actively move cations or anions into or out of the cell. When using these opsins as a tool to control neuronal...plasmids that use light to control a protein or cellular activity. For example, search "Cdc42", "transcription...

  20. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    ... is technically cell-permeable, its use in live cells can sometimes result in cellular toxicity due to... Dyes that can pass through cell membranes can prove useful for live-cell imaging. Experimental context...Additionally, cell permeability is a critical factor, as it influences the dye's ability to enter cells without...enabled the visualization of both organic tissues and cell cultures, opening biological interiors to many inquisitive...interfere with protein expression or function in live cells. Fluorophore properties: The specific properties...context of the experiment, such as the presence of cells or specific proteins, can influence the choice of...ligands is unaffected by additional protein from cells under certain conditions, which is an important ...
Showing: 221 - 240 of 813 results