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We narrowed to 710 results for: tat

Showing: 261 - 280 of 710 results

  1. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    Published
    Feb. 4, 2025, 2:15 p.m.
    ... 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...

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

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

  3. Truncated gRNAs for Regulating Gene Expression

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

  4. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

    Type
    Blog Post
    Published
    June 12, 2014, 3:30 p.m.
    ...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...

  5. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    Published
    July 31, 2020, 12:30 p.m.
    ...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...

  6. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    Published
    Nov. 29, 2022, 2:15 p.m.
    ... 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...

  7. DNA Purification Without a Kit

    Type
    Blog Post
    Published
    Feb. 11, 2020, 2:15 p.m.
    ...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...

  8. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Oct. 14, 2015, 2:30 p.m.
    ...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...

  9. A Career in Grants: How To Become a Grant Professional

    Type
    Blog Post
    Published
    Aug. 20, 2015, 2:30 p.m.
    ...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.  ...

  10. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    Published
    Aug. 20, 2020, 2:30 p.m.
    ...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...

  11. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    Published
    April 16, 2020, 1 p.m.
    ...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...

  12. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    Published
    May 8, 2018, 1 p.m.
    ...  ICE uses Sanger sequencing data to produce quantitative, NGS-quality analysis of CRISPR editing - enabling...For a multiplex sample, ICE includes visual representations of all detected edit types in the sample. The...complete, a new screen appears with a graphical representation of the results and a list of the analyzed samples...in your edited population and their relative representation in the edited pool. The black vertical dotted...

  13. Transferable Skills Guide: External Collaborations

    Type
    Blog Post
    Published
    April 19, 2018, 1:25 p.m.
    ..., allows us to display high-quality maps and annotations without us having to invest too much of our internal...partner, connect the relevant teams, coordinate implementation, and now must focus on managing the established...I personally felt that I had to overcome some hesitation to ‘approach strangers’ and learn how to communicate...maintaining useful notes that remind me about the current state of a partnership. Every collaboration and partnership...

  14. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

    Type
    Blog Post
    Published
    Aug. 13, 2019, 12:58 p.m.
    ...have since been initiated and today thousands of mutations in genes affecting diverse biological processes—not...deactivate specific genes are needed. Targeted mutation has been demonstrated in zebrafish using zinc-...depending on whether a gene is deactivated via mutation or morpholino. This observation unlocked novel... been tracked using microscopy and advanced computational approaches (Keller et al., 2008). This microscopy-based...

  15. Bioinformatics at Addgene

    Type
    Blog Post
    Published
    July 14, 2022, 1:15 p.m.
    ...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...

  16. Viral Production at Addgene

    Type
    Blog Post
    Published
    Oct. 24, 2023, 1:15 p.m.
    ...preparation by precipitation in 10% polyethylene glycol (PEG) followed by centrifugation. Precipitated pellets...i.e., genome-containing) AAV particles within representative vector preparations was quantified with electron...for viral production are propagated in the endA-mutated NEB Stable strain of E. coli. In addition, plasmids...

  17. Hot Plasmids: Fall 2024

    Type
    Blog Post
    Published
    Nov. 21, 2024, 1:30 p.m.
    ...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...

  18. Plasmids 101: Control Plasmids

    Type
    Blog Post
    Published
    April 9, 2015, 3:29 p.m.
    ... constant variables that enable the correct interpretation of the effect of the independent variable you...controlled for in order to eliminate any alternative interpretation of the results. Typically, plasmid-based experiments...non-targeting control is critical to the correct interpretation of the results because it provides an important...etc, but outside constraints may impose some limitations and you should be cognizant of these when interpreting...

  19. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    Published
    April 3, 2014, 8:05 p.m.
    ...sense OR antisense transcript depending on its orientation to your gene. T7lac High levels of gene expression...sense OR antisense transcript depending on its orientation to your gene. araBAD General expression Promoter... promoter with somewhat weak expression. lacIq mutation increases expression of the repressor 10x, thus...sense OR antisense transcript depending on its orientation to your gene Although this list is a great...

  20. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    Published
    May 5, 2020, 1:15 p.m.
    ...post was contributed by Shravanti Suresh from Iowa State University. Since its appearance, SARS-CoV-2 has... fluorescence can be observed under either no excitation light, UV light or LED blue light.  Highlights...to our guest bloggers Shravanti Surest from Iowa State University. Shravanti Suresh is a PhD candidate ...CRISPR/Cas immune systems in the Sashital Lab, Iowa State University. She is actively interested in science...
Showing: 261 - 280 of 710 results