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Showing: 561 - 570 of 1073 results
  1. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    ...Acr) proteins are phage-derived small protein inhibitors of CRISPR-Cas systems that help phages evade ...the best time to shut off Cas activity? Using inhibitor timing experiments, the Doudna lab found that ...also be used to reduce off-target edits of base editors, which convert a nucleotide base pair to another... means they can be delivered in vivo using AAV vectors. Having a way to shut off CRISPR-Cas systems prevents...Maxwell KL, Doudna JA (2017) A Broad-Spectrum Inhibitor of CRISPR-Cas9. Cell 170:1224-1233.e15 . https...Papayannopoulou T, Lieber A (2018) HDAd5/35++ Adenovirus Vector Expressing Anti-CRISPR Peptides Decreases CRISPR...
  2. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    ...biosensor affinity for calcium.”  Primary Vector Type The vector type should describe how your plasmid ...indicate type in “Vector Type Other” column. For example, you may enter “donor template vector”. Cloning...plasmids containing the ccdB gene, such as Gateway vectors, choose the ccdB Survival strain. For plasmids ...Distribute pending QC. The name of the Principal Investigator and Organization where your plasmids were first...
  3. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    ...ask the depositing laboratory to review the discrepancies. If the depositing laboratory confirms that these...reference sequence or a sequence provided by the depositor. Depositing labs often provide insert sequences... actin promoter, and rabbit beta-globin splice acceptor, contains sequence that is over 80% GC. Some IRES...information about our sequencing results in the Depositor Comments section at the bottom of the plasmid ...hope to make available as part of our OpenBio Repository sometime soon. Stay posted!     References ...
  4. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...My lab's vector of choice is AAV, with nearly every experiment requiring AAV. Before joining my lab, ... a heated stir plate to dissolve and requires monitoring so that the solution doesn’t get too hot because...approach gives me a bit of flexibility since I can store the cell lysates at -80 °C for up to 6 months before... completed in one long day, or the virus can be stored at 4 °C and buffer exchanged the next day. Refer...Production of recombinant adeno-associated viral vectors for in vitro and in vivo use. Current protocols...tool for gene expression in mammals Use the PEI calculator for planning AAV packaging transfections Find...
  5. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    ...-3.13. To test xCas9 3.0-3.13, Hu et al. first restored catalytic residues D10 and H840. To ensure that...editing Hu et al. also engineered transcriptional activators using the xCas9 scaffold. With an NGG PAM, xCas9..., confirming the flexibility of xCas9 3.7. Base editor variants with xCas9 are similarly flexible. Like...xCas9 3.7 fused to third generation cytidine base editor architecture (BE3) slightly improves NGG PAM editing...a smaller increase in NGA editing. Adenine base editor xCas9 3.7-ABE also follows the same pattern, with...Plasmids for Mammalian Systems Learn about CRISPR History ...
  6. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    ...Enhancers are cis regulatory DNA sequences that, when bound by transcription factors, can increase a gene...CAPTURE (CRISPR Affinity Purification in situ of Regulatory Elements) method. The original CAPTURE method...2020). CRISPR Affinity Purification in situ of Regulatory Elements: The original CAPTURE method The original... three components: dCas9 with an added biotin acceptor site BirA, a biotin ligase a gRNA(s) targeting...patterns in chromatin interactions. The chromatin story told by CAPTURE 2.0 is important, but it’s not the...
  7. Academic vs. Industry Postdocs

    Type
    Blog Post
    ... post was contributed by Laura Desrochers, a postdoctoral scientist at AstraZeneca. Why I chose an industry...time working on something extremely risky or exploratory. Getting a job afterwards: Fairly or unfairly...an industry setting. Scientific community and mentorship: As an industry postdoc, you might find yourself...project’s or a team’s, it is possible that your mentor (and even their department) could be laid off while...Desrochers! Laura Desrochers is currently a Postdoctoral Scientist at AstraZeneca. She has a background...blog posts here Find 9 tips for a successful postdoctoral experience Read Joanne Kamen's 8 tactics for...
  8. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    ...localization, dynamics, and behavior of a POI. As an investigator, you want a fusion tag to be inert, to not produce...has gone into making FPs monomeric, yet many investigators remain ignorant of FP dimerization. Equally ... protein fusions Transmembrane proteins, i.e. receptors and transporters, are integrated into lipid bilayers...spiderweb-like pattern of tubules can be grossly distorted into dense stacked membranes termed Organized ...from Oregon Health Sciences University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz... His interests include the quality control of secretory proteins in the Endoplasmic Reticulum and live...
  9. Genetic Code Expansion

    Type
    Collection
    ...amino acid incorporation. ID Plasmid Gene/Insert Vector Type PI Return to top Bacterial Strains Browse ...sites changes to UAG, RF1 function removed, MutS restored so decreased mutation rate George Church 68306...tolC, and tyrU gene replaced by GentR) and release factor 1. Expresses archaeal MjTyrRS/tRNA pair instead... V. W. (2021). Genetic Code Expansion: A Brief History and Perspective . Biochemistry, 60 (46), 3455–3469...
  10. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    ...available by both the Jacob Corn and Alex Schier laboratories. Delivering Cas9-gRNA ribonucleoproteins  An...This technique utilizes Cas9 proteins harboring receptor ligands (C in the figure above), which result ... potential complications, there are additional factors to take into account with these types of experiments...and purification, which will require additional laboratory reagents, equipment, and expertise. Final thoughts...production of the Cas9 protein and a gRNA in your laboratory, once these have been acquired, genomic editing...Central PMCID: PMC4289409. 3. Rouet, Romain, et al. "Receptor-Mediated Delivery of CRISPR-Cas9 Endonuclease ...
Showing: 561 - 570 of 1073 results