We narrowed to 867 results for: TOR
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TypeBlog PostPublishedMay 19, 2020, 1:15 p.m....cerevisiae expression vectors are available at Addgene. A set of mammalian expression vectors containing various...expression using either TEF1 or GAL1 promoters. These vectors are intended for the high level expression and ...spike protein) cloned into a CMV driven expression vector are also available. These contain no tags and are... genes in E. coli and S. cerevisiae expression vectors in late March. The first set of 383 constructs ...status we call “Archived.” This status allows us to store plasmids and make them available to scientists without...
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Behind the scenes: Addgene’s new search engine and more
TypeBlog PostPublishedDec. 12, 2019, 2:22 p.m....have perfected processes to curate the data our depositors share and enrich it with sequencing data; as ...categories — catalog, educational resources, and depositors — each with subcategories such as plasmids and...educational resources, and labs and institutions for depositors (see below). Even if the user experience is similar...scoped to plasmids shows filters for expression and vector type, while a search scoped to viral preps shows... match. For example, matches of a name in the depositor field will rank higher than matches of the same...that contains a plasmid with the catalog ID). Depositor suggestions provide shortcuts to matching PI labs... -
Why and how to start studying chromatin (with Addgene's help)
TypeBlog PostPublishedJune 12, 2018, 11:40 a.m....this list, that does not necessarily mean the repository does not have it. That said, if you know of a... don’t hesitate to start the process! Addgene depositors who study chromatin The PIs linked below are ...Chromosome Organization. These aren’t the only investigators in the chromatin field who have deposited with...Addgene website. In-exhaustive List of Addgene Depositors Who Study Chromatin Cheryl Arrowsmith James...SNF-like BAF complex to chromatin after T lymphocyte receptor signaling. Zhao K, Wang W, Rando OJ, Xue Y, Swiderek...Bystricky, Ben Martin, and Berkley Gryder. Addgene depositors or not, your time is valuable ... -
CRISPR 101: Non-Homologous End Joining
TypeBlog PostPublishedApril 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, ... -
A Deep Dive into BioBuilder
TypeBlog PostPublishedOct. 11, 2017, 1:44 p.m....textbook, a laboratory kit, and support during the academic year from remote scientist mentors and the BioBuilderClub...effectively contribute as part of a team.” As a seasoned educator, Kuldell also knew that the best days in the classroom...meetings and newsletters, pairs teams with STEM mentors, and provides a nationally recognized platform ...state-of-the-art lab embedded in LabCentral’s community of innovators that opened in August 2017. This flexible teaching...BioBuilder! Mary Tamer is a Boston-based writer, editor, and consultant whose work is focused on education... -
Kiran Musunuru on the Newest TALEN Genome-Editing System
TypeBlog PostPublishedDec. 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... -
Reproducibility for Everyone: Lessons from an Open Science Collaboration
TypeBlog PostPublishedFeb. 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... -
Addgene's Tips for Plasmid Quality Control
TypeBlog PostPublishedJan. 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 ... -
Gendered Innovations: Why Does Sex of the Cell Matter?
TypeBlog PostPublishedJuly 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... -
Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene
TypeBlog PostPublishedApril 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 ... -
Hot Plasmids and Viral Preps - January 2021
TypeBlog PostPublishedJan. 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... -
CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems
TypeBlog PostPublishedJuly 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... -
Engaging with science and society at pgEd
TypeBlog PostPublishedSept. 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... -
Overcoming the Challenges of Lentiviral Production
TypeBlog PostPublishedAug. 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 ... -
Quickest Way to Deposit Plasmids: The Deposit Spreadsheet
TypeBlog PostPublishedMarch 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... -
Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR
TypeBlog PostPublishedJuly 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... -
Tips for a 1st time AAV user (by a Rookie AAV user)
TypeBlog PostPublishedOct. 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... -
xCas9: Engineering a CRISPR Variant with PAM Flexibility
TypeBlog PostPublishedMarch 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 ... -
When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells
TypeBlog PostPublishedApril 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... -
Academic vs. Industry Postdocs
TypeBlog PostPublishedNov. 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...