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We narrowed to 820 results for: cell;

Showing: 441 - 460 of 820 results

  1. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ... into a mouse embryonic stem cell (ESC) line, enrich and select for cells that had successfully incorporated... when it gets exciting – the cell sends out an “SOS” signal and cellular repair mechanisms rush in to ...Cas-Mediated Genome Engineering. Cell 154:1370–1379 . https://doi.org/10.1016/j.cell.2013.08.022 Yoon Y, Wang ...Cas-Mediated Genome Engineering. Cell 153:910–918 . https://doi.org/10.1016/j.cell.2013.04.025 Zhang X, Liang...develop into chimeric mice (with a fraction of the cells in the adult mice derived from the engineered ESCs...Humanization (25-kbp) Created Using Embryonic Stem Cell/Blastocyst and CRISPR/Zygote Injection Approaches...

  2. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    ... Guide RNA and Target DNA. Cell 156:935–949. https://doi.org/10.1016/j.cell.2014.02.001  Ran FA, Cong ... Delivery by AAV Vectors. Cell 181:136–150 . https://doi.org/10.1016/j.cell.2020.03.023 Additional Resources...acids) displayed cleavage activity in mammalian cells. SaCas9 produced indels at a similar efficiency ...package each piece into an individual AAV. When a cell is transduced by both AAVs, the full length gene...SpCas9 which had modest editing rates in HEK-293T cells when compared to the full-length SpCas9. In 2016... allows cleavage of HBB and CCR5 genes in human cells using compact expression cassettes. Sci Rep 5: ....

  3. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    ...interest live within a cell,” “Does this disease change the architecture of my cells,” or “How does this ...this mutation impact the types of cells found in my tissue.” It is based on the same principles as other ... visualize a target of interest within an intact cell, tissue, or, in some cases, a whole organism. But...joined to fluorescent molecules are applied to a cell or tissue sample where they bind to their targets...to visualize a target within an intact sample, a cell in the case of ICC or a tissue in the case of IHC...IF), is an immunoassay that brings to light the cellular world. The technique allows you to ask questions...

  4. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    ...construct and then introduce both plasmids into the cell. The edit in the chromosome can now take place. .... After editing, pTargetF can be cured from the cell by inducing expression of the sgRNA that targets ...all the edits you want, cure pCas9 by growing the cells at 37 °C.     The new pEcCas/pEcgRNA system We ... to take place. Then plasmids are cured from the cell. Steps 1-1 and 1-2 can be done simultaneously. Image...tools for faster construction or reprogramming of cells to understand the genetic and biochemical mechanisms...phenotypes and to design a new generation of engineered cells for green chemistry and biomedical applications.... Chinese Academy of Sciences (CAS) Center for Excellence in Molecular Plant Sciences in 2019. She joined...

  5. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ... want to make sure your cells have first reached an appropriate density. Cells in the exponential growth...collecting dead cells, and, conversely, you cannot induce too early as there are not enough cells in the culture...host protein, using up valuable resources in the cell thus leading to slowed growth. Additionally, some...such as TB or 2XYT is optimal because of the high cell-density they support; however, minimal media supplemented...

  6. Keeping Up With CRISPR/Cas9

    Type
    Blog Post
    ...RNA-Mediated Gene Targeting in One-Cell Embryos. Niu Y, et al. Cell. 2014 January 30. (Pubmed) Programmable...correct disease mutations, both in mice and in human cells. But that important advance was quickly followed...

  7. Plasmids 101: Gram Negative and Positive Bacteria

    Type
    Blog Post
    ...negative bacteria is due to differences in cell wall composition. Cell wall structure Gram positive bacteria...complex within the peptidoglycan layer, staining the cell purple. When the alcohol is added, it dehydrates...Silhavy, TJ., Kahne, D. & Walker S. The Bacterial Cell Envelope. Cold Spring Harb Perspect Biol. 2(5) a000414...

  8. Water Bath Protocol

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

  9. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    ...editing systems by manipulating cellular determinants of editing outcomes. Cell, 184(22), 5635-5652.e29. https...efficient prime editors. Cell, 186(18), 3983-4002.e26. https://doi.org/10.1016/j.cell.2023.07.039 Lin, Q.,...correct only a minority of those variants in most cell types. But prime editing, a CRISPR technique developed...unedited DNA. The heteroduplex is resolved by the cell’s mismatch repair system, and more recent prime editing...editing demonstrated ~20-50% efficiency in HEK293T cells with 1-10% indels. Since then, further innovations...type of edit, genomic context, delivery strategy, cell type, and goal of the experiment can all impose ...nicking the non-edited strand, they can cause the cell to remake that strand using the edited strand as...

  10. What's New in CRISPR - May 2019

    Type
    Blog Post
    ...genome-wide deamination of RNA cytosines in human cells. Thus, they engineered two CBE variants with mutations...RNA edits by >390 fold and >3,800 fold in human cells with no significant reduction in editing efficiency...mice and in zygotes. Read the paper in Molecular Cell Find the Nme2Cas9 All-in-One AAV constructs CRISPR...

  11. Let There Be LITE Plasmids

    Type
    Blog Post
    ... vectors and genetically target them to specific cell populations. They demonstrated use of the LITEs ...non-invasively, its use is not restricted to cultured cells but extends to freely moving animals, as established...no means limited to particular model organisms or cell types. “This tool is really flexible and modular...novel mode of optogenetic control of endogenous cellular processes." The LITE system can now be applied...

  12. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

    Type
    Blog Post
    ...Control of Gene Expression. Cell 152:1173–1183 . https://doi.org/10.1016/j.cell.2013.02.022 Sajwan S, Mannervik...and Fluorescence Imaging. Cell 159:635–646 . https://doi.org/10.1016/j.cell.2014.09.039 Zhou H, Liu J,...from when looking to activate genes across diverse cell lines.   dCas9-VP64 Description CRISPR activation...large and are not expressed consistently throughout cells. VPR Description VPR fuses a tripartite complex ...activation levels than SAM in some loci across Drosophila cell lines (Sajwan et al., 2019). SPH SPH combines components...

  13. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    ...Inhibitor of CRISPR-Cas9. Cell 170:1224-1233.e15 . https://doi.org/10.1016/j.cell.2017.07.037 Li C, Psatha...Off-Switches for CRISPR-Cas9. Cell 167:1829-1838.e9 . https://doi.org/10.1016/j.cell.2016.11.017 Shin J, Jiang...concern is that the longer Cas remains active in a cell, the greater chances there are for off-target edits...within the first six hours. When added to human cells six hours after the introduction of Cas9 RNPs, AcrIIA4...CRISPR/Cas9 Toxicity in Human Hematopoietic Stem Cells. Molecular Therapy - Methods & Clinical Development...

  14. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    ...Single-Domain Camelid Antibodies. Cell 181:1004–1015.e15 . https://doi.org/10.1016/j.cell.2020.04.031 Zimmermann ...coronaviruses and enable the virus to enter host cells. To do this, they injected the spike protein from...camelid antibodies that bind to the spike proteins in cell culture, and inhibit these viruses.  But llama ...hidden in the molecular crevices proteins on the cell surface. Figure 1: Comparison of common antibody...blog Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification RANbodies: Reporter...

  15. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ... genetic loci within a single cell. One might imagine transfecting cells with both the nuclease-positive...nuclease-competent Cas9, obviating the need to generate new cell lines or transgenic animals expressing dCas9 as ...reliance on orthogonal Cas9 species. Within a single cell, full-length guides targeting one set of genetic...knockin mice for genome editing and cancer modeling." Cell 159.2 (2014): 440-455. PubMed PMID: 25263330. PubMed...

  16. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ... for Mapping Multiprotein Complexes in Human Cells.” Cell Rep. 13: 621-633. PMID: 26456817. Find plasmids...Chromosomes). Using their PITCh plasmids, GFP knock-in cell lines can be created in about a month and a half...locus-specific gRNAs Step 4: Select for puromycin resistant cells Step 5: PCR amplify and sequence the locus to verify...crispr.mit.edu. They tested their system in HEK293 cells, integrating the GFP-Puro cassette into the FBL ...

  17. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    ...fluorescence-activated cell sorting (FACS). This is not possible in plants outside of limited, single cell protoplast...genes known to be essential for cell growth. Together, this excellent work highlights the importance of...visually identified subpopulation. For traditional cell culture, enriching your active RNAi subpopulation...where he investigated how plants achieve polarized cell growth in the lab of Luis Vidali. For his next adventure...Screens for Effective Gene Silencing in Plants. Plant Cell 25:1507–1522 . https://doi.org/10.1105/tpc.113.112235...Augmin in Phragmoplast Microtubule Generation. Plant Cell 24:1478–1493 . https://doi.org/10.1105/tpc.112.098509...exquisitely and robustly self-organize at the cellular and multicellular scale.     References Bezanilla M, Perroud...

  18. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...at lower copy numbers within bacterial cells, and bacterial cells with multimers grow more slowly compared...stable and more prone to being lost in bacterial cells compared to monomers, although this can be prevented...increase the frequency of base substitution in human cells. J Biol Chem. 2011 Mar 25;286(12):10017-26. doi:...plasmids leading to the formation of plasmid multimers. Cell. 1976 Dec;9(4 PT 2):707-16. doi: 10.1016/0092-8674...supercoiled plasmid DNA concatemers in mammalian cells. J Gene Med. 2009 May;11(5):444-53. doi: 10.1002...essential for plasmid monomerization and stability. Cell. 1984 Apr;36(4):1097-103. doi: 10.1016/0092-8674...

  19. CRISPR Plasmids - C. elegans

    Type
    Collection
    ...sequence. DSBs are preferentially repaired in the cell by non-homologous end joining (NHEJ), a mechanism...PAM site. When a repair template is present, the cell may repair a DSB using homology-directed repair ...Cas protein and a target-specific gRNA in the same cell at the same time. Single plasmids containing both...

  20. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    ...miRNA and piRNA can be found endogenously in many cell types, while shRNA and siRNA are synthetic constructs...methods. shRNA processing Following delivery into cells using viral vectors or plasmids, shRNAs hijack the...shRNAs is to generate stable knockout or knockdown cell lines. In fact, it was the go-to choice for many...shRNAs are a quick way to create stable knockdown cells lines, and you can usually find validated shRNA ...Resources References Bartel, D. P. (2004). MicroRNAs. Cell, 116(2), 281–297. https://doi.org/10.1016/s0092-...degradation of RNA-induced silencing complex. Molecular Cell, 82(1), 30–43. https://doi.org/10.1016/j.molcel.2021.11.026...
Showing: 441 - 460 of 820 results