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We narrowed to 844 results for: Tat

Showing: 381 - 400 of 844 results
  1. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

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    Blog Post
    ...consequences of that change. For example, “Mutated aspartate 123 to Lysine (D123K), lowers biosensor affinity...indicated in the “Species Other” column. Relevant Mutations Indicate the amino acid change as well as any...accept sequence files in any format. Distribution status for your plasmids: Hold for Publication or Distribute...
  2. Engaging with science and society at pgEd

    Type
    Blog Post
    ...understanding of how our bodies work, new treatments for debilitating diseases, and maybe even ways to alleviate health... broaden the conversations about genetics and facilitate dialogue between scientists and communities. ...With our focus on engaging the community and facilitating dialogue, a major part of our everyday work ...jargon-filled language in academic writing and presentations. But for many people who don’t work in these...
  3. Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux

    Type
    Blog Post
    ...turn. The first is the passing and upcoming implementation of the Federal Labor Standards Act (FLSA) which... in salary, particularly given that this has precipitated increases to the NIH recommended pay scale –...and opportunities for us all.  For example, implementation and compliance for the FLSA changes, including...challenging, but will be aided by the development and implementation of new strategies designed to cope with current...
  4. Gaining Leadership Skills Volunteering at a Professional Organization

    Type
    Blog Post
    ... NPA states that Postdoctoral scholars should have the skills and techniques needed to facilitate effective... Chair automatically as it was an understood expectation of any Vice-Chair. Following an official assessment...leader in the NPA, I had to speak as an NPA representative and speak the NPA’s stance. I had to be professional...
  5. The AAVantages of AAV in CRISPR Screens

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    Blog Post
    ... However, safety requirements and expression limitations may be too constraining for some experiments,...in your experiment. Figure 2: Visual representation of common AAV serotypes used for different ...that reside outside a host cell’s genome. In this state, AAV cargo can be stably expressed in non-dividing... on the Addgene Blog Overcoming the AAV Size Limitation for CRISPR Delivery Genome-wide Screening Using...
  6. Plasmids 101: Degron Tags

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    Blog Post
    ... or C terminal of a protein and can inducibly facilitate the degradation of the tagged protein. Degrons...here are all conditional, meaning that they only facilitate degradation of their fused protein under certain...through interacting ligands. After an E3 ligase facilitates ubiquitination of the Halo-tagged protein in ...HaloTag and its PROTAC are all that is needed to facilitate degradation. HaloTag is also the degron tag with...
  7. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...molecular biology and bioengineering, as it has facilitated the development of a simple and scalable means... gene expression. Its catalytic domains can be mutated to inactivate the protein’s nucleolytic capability...interrogation of gene function. However, there are limitations to a system that relies on two distinct Cas9 ...possible, this strategy suffers from several limitations. Perhaps most importantly, most Cas9 orthologs...
  8. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...approach for harvesting my AAV. I skip the PEG precipitation of the HEK293 cell culture supernatant because...pellet. My yield would be higher if I used PEG precipitation, but for my experiments, I get enough virus ...culture media, you should consider performing PEG precipitation. Instead of sonicating the cell pellet, I lyse...with this AAV lysis buffer and freeze/thaw the lystate four times in a 95% ethanol and dry ice bath to...
  9. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    ...accounting for some of the inherent inaccuracies of computational prediction models. In addition, we provide options...most promising pegRNA designs and especially facilitates the use of prime editing in pooled settings with...models (such as PRIDICT/PRIDICT2.0) can greatly facilitate prime editing experiments, thus saving you time...Switzerland), where he developed a strong passion for computational biology. His expertise includes applying machine...
  10. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

    Type
    Blog Post
    ...using the MoClo system. The paper also gives quantitative data for regulatory elements, which has not ...example, knock out a gene with simultaneous complementation, with a knockout and expression construct on...tools, double dimerization GFPs, and circular permutation GFPs. Hopefully we will have new tools for plant...doing. They look at structure, impediments, the representation of women on committees, and how the culture...
  11. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    ... knew that their study had significant statistical limitations, and expected to find only a few, very ...theory, if the DepMap analysis had an equal representation of every ancestry group, they should have found... researchers to ignore or miss. The lack of representation in commonly used cell lines led to an incorrect...
  12. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ...like guanine. This RNA editor would permit point mutations in RNA which could recapitulate or rescue known...REPAIRv1, the lab targeted 34 pathogenic G->A mutations from the ClinVar database. They successfully edited...still displays substantial off-target activity. By mutating residues in ADAR2 that interact with duplex RNA...rational mutagenesis and identified that a S375A mutation in RESCUE (named RESCUE-S) offered ~76% on-target...
  13. DNA Purification Without a Kit

    Type
    Blog Post
    ...washed, and either eluted from a fixed surface or precipitated, dried, and then rehydrated. There are some ...which causes the proteins and genomic DNA to precipitate. Plasmid DNA is free in solution. Proteins and...remaining proteins and RNase. Finally, DNA is precipitated with alcohol and resuspended in TE buffer. After... the National Academy of Sciences of the United States of America, 76 2, 615-9 . https://doi.org/10.1073...
  14. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    ...systems. A putative new class 2 nuclease, Cpf1 (CRISPR from Prevotella and Francisella), was annotated in several...sophisticated genome-wide analysis. At many of its computationally predicted off-target sites, Cpf1 does not mediate...learn about CRISPR biology. Later work, like the adaptation of Cas13 to RNA targeting and RNA editing, has...
  15. A Career in Grants: How To Become a Grant Professional

    Type
    Blog Post
    ...compliance with the grantor’s requirements as well as state and federal regulations. A grant professional’s ...no surprise that the U.S. Bureau of Labor and Statistics shows that employment of grant writers is expected...consider consulting for a particular grant by another state. With the blessing of my employer, I took on my ...application writing and development, project implementation, program evaluation and grant management.  ...
  16. Bioinformatics at Addgene

    Type
    Blog Post
    ...data pipeline is to perform several sequential computational tasks for which existing script(s) have been...  We started at the usual recommended place: documentation. We oriented ourselves with the basic concepts... the file name. We started small with a print statement and tested that the desired data was available...we manually inspected the output with a print statement at the end of the script or by opening the files...
  17. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ...These off-target sequences may be unintentionally mutated along with the desired target locus. The PAM sequence..., we will cover three ways to circumvent this limitation: 1) the use of novel S. pyogenes Cas9 variants...PAM binding specificity: SpCas9 variant Mutations (relative to SpCas9) PAM sequence D1135E...human genome. Another variant, xCas9 3.7 has 7 mutations found in the REC2, REC3, and PAM interacting domains...
  18. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    ...determine the presence and quantity of DNA, RNA or a mutation of interest. Together SHERLOCK and DETECTR demonstrate... specificity.  SHERLOCKv2 SHERLOCK had a few limitations when it was first introduced in 2017 so the lab... pre-amplification step allowing for greater quantitation without compromising sensitivity. To detect...SHERLOCK could also detect low-frequency cancer mutations from cell-free DNA fragments as well as health-related...
  19. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

    Type
    Blog Post
    ... same (forward) or in the opposite (reverse) orientation? The answers to questions like these will determine...sequences, called modules or parts. Assembly is dictated by four-base overhangs that are paired with different... the sequence, normally through the synonymous mutation of a codon. You can domesticate the sites yourself...colony PCR electrophoresis), is free of undesired mutations, and that you have the correct four-base overhangs...
  20. Hot Plasmids: Fall 2024

    Type
    Blog Post
    ...TBD domains, part of the HB domain, and several mutations to improve solubility and stability (Figure 2)...through eight stages of embryonic development. Representative images of single focal planes through the middle...in natural DNA repair pathways and spontaneous mutations.  Here, the researchers were able to target multiple...abscessus either simultaneously or stepwise, achieving mutation efficiencies of up to about 9% of colonies (102...
Showing: 381 - 400 of 844 results