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We narrowed to 863 results for: TOR;

Showing: 441 - 460 of 863 results

  1. Plasmids 101: Screens vs. Selections

    Type
    Blog Post
    Published
    Nov. 10, 2022, 2:15 p.m.
    ...define what type of factor you are looking for and what process or outcome that factor functions in. The...there is removal of known factors (such as a genes) and the loss of those factors is assessed. The two most... committed to the daunting task of finding new factors in your biological system of interest, the so-called...selected growth conditions. For screens, there is no obligatory growth advantage – that is, you are not specifically...

  2. CRISPR 101: Non-Homologous End Joining

    Type
    Blog Post
    Published
    April 16, 2015, 3:45 p.m.
    ...-introduced breaks are repaired. The following factors are required for NHEJ repair regardless of end ...XRCC4-ligase IV); together with some accessory factors (PAXX, XLF), this complex holds a pair of DNA ends...Accordingly, NHEJ has a vast toolbox of processing factors, including polymerases (Pol μ and Pol λ), nucleases... sequence (“microhomologies”). NHEJ doesn’t obligatorily introduce indels. Given the end structure of...majority of their chromosomes within a day. Another factor expected to impact repair is that the Cas9 protein... loading of Ku and normal NHEJ activity. Other factors can also influence NHEJ activity. For example, ...

  3. Kiran Musunuru on the Newest TALEN Genome-Editing System

    Type
    Blog Post
    Published
    Dec. 10, 2013, 3:13 p.m.
    ...pluripotent stem cells: a transcription activator-like effector nuclease (TALEN) kit consisting of 834 ...stem cells. Addgene: Tell me about the history. How had investigators done this kind of work before? Musunuru...Unfortunately, it is time consuming and difficult for investigators without a lot of expertise to create zinc finger...and puts it in the hands of pretty much all investigators. Our kit makes it extremely simple to design...

  4. Reproducibility for Everyone: Lessons from an Open Science Collaboration

    Type
    Blog Post
    Published
    Feb. 28, 2019, 1:34 p.m.
    ...already run several reproducibility workshops as Director of Scientific Outreach at Code Ocean and through...well received, and then when the team started brainstorming ideas for the next workshop, they realized that...workshop at the conference, but because each contributor made the materials accessible, anyone could present...and were presented at the ASPB meeting by the collaborators who could participate. Despite the group not...visibility of these materials, Sonali Roy, a postdoctoral researcher at the Noble Research Institute, ... Schwessinger for helpful conversations on the history of the group, lessons learned, and to Sonali Roy...

  5. Gendered Innovations: Why Does Sex of the Cell Matter?

    Type
    Blog Post
    Published
    July 21, 2016, 2:30 p.m.
    ...there are many factors that interact with sex, and it’s important to analyze these factors during an experiment...blogger Londa Schiebinger, PhD, Hinds Professor of History of Science, Stanford University. Sex and gender...experiment. In the case of stem cells, these factors may include cell type, the disease being treated, ...other hormonal, immunological, and environmental factors. How to include sex in experimental design Recognizing...variable in all NIH-funded research. Janine Clayton, Director of the Office of Research on Women’s Health, emphasizes...

  6. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    Published
    Jan. 14, 2016, 3:30 p.m.
    ...ITRs), hairpins, or strong terminators. Detailed instructions on other factors to consider when designing...community. This is no small task, however, at a repository that consistently receives around 200 new DNA...are sequencing from the exact same stock that we store and distribute from. Of course, your culture should...overnight cultures for glycerol stock production and storage, we grow a little extra and transfer 100 µl of ...Pages Get Plasmid Backbone Information from Our Vector Database  ...

  7. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    Published
    March 8, 2017, 3:15 p.m.
    ...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...

  8. Engaging with science and society at pgEd

    Type
    Blog Post
    Published
    Sept. 20, 2018, 12:17 p.m.
    ...This guest post was contributed by Johnny Kung, Director of New Initiatives for the Personal Genetics Education...dialogue between scientists and communities. As the director of new initiatives at pgEd, I take on a variety...education and government; advising writers and directors in Hollywood; and working directly with community...sticky ethical dilemmas to dark episodes in the history of eugenics and research misconduct. Past ethical... in genetics research, their applications and regulatory policies; keeping tabs on the conversation about...implications of their work. Johnny Kung is the Director of New Initiatives for the Personal Genetics Education...

  9. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    Published
    Aug. 7, 2018, 12:31 p.m.
    ... While lentiviral vectors are popular gene delivery tools, producing lentivirus, can pose certain challenges...systems such that it no longer requires the viral transactivator, Tat. For more information on the different...of concentration method depends of a variety of factors such as resources available, cost and downstream...determination of the packaging limit of lentiviral vectors. Hum Gene Ther. 2001 Oct 10;12(15):1893-905. PubMed...Tips for titering your lentivirus Visit the viral vector featured topic page Resources on Addgene.org ...Guide Find lentivirus plasmids Browse all viral vectors ...

  10. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    Published
    April 6, 2021, 12:30 p.m.
    ...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 ...

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

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

  12. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    Published
    Jan. 5, 2021, 2:15 p.m.
    ...engineered prime editors to expand their PAM flexibility. Find the engineered prime editors here A lentiviral...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...pubmed.ncbi.nlm.nih.gov/24390935/ Hypercompact genome editor CRISPR-Casɸ By Melina Fan The Doudna Lab adds to...discovery of CRISPR-Casɸ, a hypercompact genome editor from the Biggiephage clade of huge phages. In this... Scientists looking for more compact genome editors due to delivery constraints and scientists seeking...

  13. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    Published
    Oct. 23, 2018, 11:49 a.m.
    ...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...

  14. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    Published
    March 28, 2018, 6:52 p.m.
    ...-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 ...

  15. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    Published
    July 2, 2020, 12:40 p.m.
    ...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...

  16. Academic vs. Industry Postdocs

    Type
    Blog Post
    Published
    Nov. 13, 2018, 1:22 p.m.
    ... 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...

  17. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    Published
    April 19, 2016, 2:30 p.m.
    ...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...

  18. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    Published
    Aug. 13, 2024, 1:15 p.m.
    ...production is Komagataella phaffii (a.k.a  Pichia pastoris) because it secretes fewer of its own proteins...using a desalting column or an ultrafiltration concentrator. As the name suggests, you may also concentrate... to their parental hybridomas. Aggregation and storage Aggregation of antibodies may impact their quality...testing. An aliquot of our recombinant antibodies is stored at 37 °C for a period of two weeks. After this ...we are also cautious during the aliquoting and storage process to prevent dropped tubes. Now that your...Varley P, Rockberg J, Chotteau V. Small-scale bioreactor supports high density HEK293 cell perfusion culture...

  19. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog Post
    Published
    Dec. 22, 2022, 2:15 p.m.
    ... (NHEJ) and alternative end joining (alt-EJ). Inhibitors to essential components of both pathways have...events (Yang et al., Int J Mol Sci). Adding these inhibitors to your culture media prior to and after Cas9...introduction increases HDR, just make sure to remove the inhibitors within a day or two to not affect the viability... one step further, you can inhibit cell cycle regulatory proteins responsible for the transition out of...to fuse an HR protein to the nuclease. Early HR factors fused to Cas9 have been shown to increase HDR frequency...committing a break to performing HR before other factors were loaded onto the break, potentially shuttling...

  20. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    Published
    Sept. 6, 2018, 12:02 p.m.
    ...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: 441 - 460 of 863 results