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We narrowed to 657 results for: cell;

Showing: 361 - 380 of 657 results

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

    Type
    Blog Post
    Published
    April 13, 2021, 1:15 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Feb. 10, 2015, 3:06 p.m.
    ... 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...

  3. Keeping Up With CRISPR/Cas9

    Type
    Blog Post
    Published
    March 11, 2014, 5:55 p.m.
    ...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...

  4. Plasmids 101: Gram Negative and Positive Bacteria

    Type
    Blog Post
    Published
    Aug. 2, 2022, 1:15 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Jan. 13, 2025, 9:49 p.m.
    ...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...

  6. What's New in CRISPR - May 2019

    Type
    Blog Post
    Published
    May 28, 2019, 12:49 p.m.
    ...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...

  7. Let There Be LITE Plasmids

    Type
    Blog Post
    Published
    Nov. 21, 2013, 3:56 p.m.
    ... 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...

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

    Type
    Blog Post
    Published
    Oct. 6, 2020, 1:15 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    July 23, 2020, 1:20 p.m.
    ...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...

  10. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    Published
    June 18, 2020, 1:15 p.m.
    ...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...

  11. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    Published
    Jan. 10, 2017, 3:37 p.m.
    ... 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...

  12. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    Published
    Feb. 23, 2016, 3:30 p.m.
    ... 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 ...

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

    Type
    Blog Post
    Published
    Oct. 27, 2020, 1:15 p.m.
    ...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...

  14. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    Published
    Jan. 10, 2023, 2:15 p.m.
    ...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...

  15. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    Published
    June 17, 2025, 1:15 p.m.
    ...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...

  16. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    Published
    Nov. 7, 2014, 2:56 p.m.
    ...degraded, cell still can methylate DNA   hsdS(rk-,mk-) Unmethylated DNA not degraded, cell cannot methylate... gal Mutation in galactose metabolism pathway Cells cannot grow on galactose only gyrA, gyrA96 DNA...permease inactivated, lactose cannot be taken up by cell mcrA, mcrBC Inactivation of pathway that cleaves...gene that maintains the plasmids at ~250 copies per cell.  F- ∆lac169 rpoS(Am) robA1 creC510 hsdR514 endA...rpsL20 xyl-5 leu mtl-1 Stbl4   Electrocompetent cells for cloning and storage of unstable DNA and lentiviral...

  17. New and Upcoming Viral Vectors - May 2020

    Type
    Blog Post
    Published
    May 28, 2020, 1:15 p.m.
    ... results, but also confirm expression in target cells. While the various AAV serotypes can be used to ...Chemogenetics offers noninvasive methods for manipulating cell activity, particularly in the brain, and holds promise... systems such as Cre-lox allow for more accurate cell and tissue targeting. While promoters and AAV serotypes...allowing you to further restrict gene activation to cells expressing both Cre and GFP. The GFP dependent FLPo...

  18. Plasmids 101: Inducible Promoters

    Type
    Blog Post
    Published
    Jan. 18, 2018, 2:34 p.m.
    ...stimulate AraC binding to I1 and I2 sites. Supplementing cell growth media with glucose decreases cAMP and represses... Because transcription machinery differs between cell types or organisms, promoters are similarly variable...variable. Bacterial promoters only work in prokaryotic cells and typically only in the same or closely related... were derived. Similarly, the various eukaryotic cell types (mammalian, yeast, plants, etc.) require unique...

  19. A Novice's Perspective on Neuroscience - SfN2017

    Type
    Blog Post
    Published
    Dec. 15, 2017, 2:07 p.m.
    ...aiming to dedifferentiate mouse brain cells into neural stem cells. I was quite unsuccesful (although I...versatile means of delivering genes to many different cell types with minimal immunological effects. AAVs are... used in experiments targeting genes to specific cell types in the brain, mapping neuronal networks, and...

  20. Antibodies 101: ChIP

    Type
    Blog Post
    Published
    March 10, 2022, 3:25 p.m.
    ... Process ChIP can be done for either tissues or cells. The process is very similar for both, though tissue...fragmenting the DNA, spin down the samples to remove cell debris, which will collect in a pellet. The supernatant..., these are mostly fairly standard molecular and cell biology techniques. What’s all the fuss? Well, ChIP...will also have to be optimized for both tissue or cell type, amount of protein, and desired fragment length...
Showing: 361 - 380 of 657 results