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We narrowed to 197 results for: Anti-CRISPR

Showing: 161 - 180 of 197 results
  1. SunTag and Fluorescent Imaging

    Type
    Blog Post
    Published
    March 28, 2017, 2:30 p.m.
    ...fluorescence, SunTag can also be used to improve CRISPR-based activation of target genes, but we’ll save...GCN4. GCN4 recruits GFP fused to the cognate scFV antibody, which is expressed from a separate plasmid. This...
  2. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    Published
    Jan. 16, 2018, 3:20 p.m.
    ...translation in living single cells. However, before CRISPR-Cas9 these techniques were quite tedious and, even...are added to a cellular sample. Both antibodies have a short antibody specific DNA strand added to them....PLA where one antibody was directed toward a protein of interest but the second antibody was directed ...most common approach. With a good and specific antibody and the proper microscope setup we can observe...interactions in situ. In the standard PLA assay, primary antibodies directed towards two endogenous proteins of interests...amplification could only take place if the two antibodies recognized an endogenous protein of interest ...proteins in situ. Now FUNCAT-PLA provides both quantitative and spatio-temporal information about the turnover...
  3. Plasmids 101: NGS Quality Control for Pooled Libraries

    Type
    Blog Post
    Published
    Oct. 26, 2017, 1:59 p.m.
    ...Sequencing to Validate AAV Preps Learn More about Pooled CRISPR Libraries Resources on Addgene.org Browse Pooled... from both. These products are then purified, quantified, and sent for sequencing. Analyzing your sequencing...
  4. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    Published
    June 27, 2019, 12:25 p.m.
    ...or preparation, and deliver CRISPR components for gene disruption or CRISPRi. While designed to study the...can be both sorted using FACS and isolated with antibiotic selection. The team tested this system in vitro...the addition of standards, the assay is semi-quantitative and can approximate mRNA concentration within...host-range of the plasmids and suitability of various antibiotic resistance markers, promoters, and inducible ...microbiome responds to infection, dysbiosis, and antibiotic treatment. To facilitate these studies, Pamela...
  5. How Dry Ice Affects Viral Vector Sample pH & How to Avoid It

    Type
    Blog Post
    Published
    June 20, 2017, 2:30 p.m.
    ...Service Tips for 1st Time CRIPSR User (By a 1st Time CRISPR User) Tips for Titering Your Lentivirus Preps ... (Fig. 1). However, once thawed, there was a substantial reduction (from 7.4 to 5.0) in the pH of samples...that dry ice storage affected the performance of antibodies, because of effects of pH on protein stability...infectivity of the virus. However, there was a substantial reduction in infectivity after 3 weeks of exposure...
  6. All in a Twist: dsRNA

    Type
    Blog Post
    Published
    Feb. 2, 2023, 2:15 p.m.
    ...021-00430-1 More resources on the Addgene blog CRISPR 101: RNA Editing with Cas13  RNA Interference ...This pathway has been clinically exploited for antiviral purposes similar to that of vaccines. If you know...is certainly a double-edged sword – balancing antiviral benefits with the inflammatory response associated...balancing the amount of siRNA to be delivered or anticipating this response are important considerations when...
  7. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    Published
    March 15, 2016, 2:30 p.m.
    ... Lentivirus FAQ Learn How Lentivirus is Used in CRISPR Genome-Wide Screening Learn How Lentivirus Has ...viral prep, treated with the antibiotic, and colony-forming units quantified. While these methods tend to...of a gene carried on the transfer plasmid or quantifying the number of viral copies that have integrated...commercially available p24 ELISA kits that can quantify p24 levels in a matter of hours. A major drawback...particle. Consequently, titers based off of p24 quantification tend to be overestimates since they can include...viral RNA is first converted to cDNA and then quantified using qPCR primers targeting specific viral components... components such as LTRs, gag, WPRE, antibiotic resistance-genes, or the transgene itself. Many researchers...
  8. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    Published
    Sept. 15, 2020, 1:15 p.m.
    ... comes in. Let’s take a look at All_in_one_CRISPR/Cas9_LacZ, a CRISPR gRNA plasmid from Lynne Postovit...have gained or lost a specific gene survive (ex: antibiotic resistance marker). In a previous blog post, ... your plasmid backbone, as it often relies on antibiotic resistance. But what about identifying clones...
  9. Learning Technical Skills at a Lab Central Workshop

    Type
    Blog Post
    Published
    Feb. 20, 2018, 2:30 p.m.
    ...Lab Management Software for Biology Learn about CRISPR Pooled Libraries Resources at Addgene.org Find...Therapeutics, a biotechnology company that uses neoantigen biology to treat cancer. My partner for the day...
  10. K. phaffii: Rising to the Occasion

    Type
    Blog Post
    Published
    April 4, 2023, 1 p.m.
    ...Resources on Addgene.org  Yeast-mediated cloning CRISPR plasmids: Yeast Yeast Synthetic Biology Plasmids...of glycoproteins for human therapies, such as antibodies, is possible in K. phaffii since it produces ...
  11. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    Published
    June 29, 2017, 1:06 p.m.
    ...are commonly used for Golden Gate cloning or for CRISPR gRNA cloning. With this more functional display... a variety of common plasmid features such as antibiotic resistance genes, selectable markers, promoters...
  12. A Novice's Perspective on Neuroscience - SfN2017

    Type
    Blog Post
    Published
    Dec. 15, 2017, 2:07 p.m.
    ...interactions with Alzheimer's, I found others using CRISPR to generate new knockout mice, using adenoviral...There were, of course, many companies selling antibodies, microscopes, and dissection tools, but there...
  13. Early Career Researcher Toolbox: Finding Relevant Papers

    Type
    Blog Post
    Published
    Jan. 9, 2020, 1:45 p.m.
    ...including a bot for biosensor papers, microbe articles, CRISPR papers, and more. Saving papers There isn’t technically...about and jumping into the discussion! Semantic Scholar Semantic Scholar is an AI-backed literature search...the Allen Institute for Artificial Intelligence. Semantic Scholar analyzes publications and figures using...more relevant to your search terms. Key features Semantic Scholar has several ways to focus your search ...at the top of this filter. A unique feature of Semantic Scholar search results are the topic pages, which...displayed. Saving papers Papers can be saved to a Semantic Scholar library. You can add hashtags to library...
  14. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    Published
    May 30, 2023, 1:15 p.m.
    ...transient gene expression in a particular cell type to CRISPR genome editing, optogenetics, and chemogenetics...specificity; this is particularly helpful when wanting to target specific neuron subtypes, since AAVs ..."Development and optimization of a real-time quantitative PCR-based method for the titration of AAV-2 ...
  15. Developing a cold tolerant E. coli using specialized chaperones

    Type
    Blog Post
    Published
    Jan. 28, 2020, 2:35 p.m.
    ...temperature conditions, thereby enabling efficient CRISPR-Cas mediated editing for psychrophilic systems ...activity of these enzymes implies that even a minute quantity can significantly reduce the operational time ...
  16. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    Published
    May 24, 2018, 1:20 p.m.
    ...Biology with BioBuilder 3D Models for learning about CRISPR Resources at Addgene.org Find plasmids from the...riboprobe binding (via hybridization) specific antibodies that detect the mRNA-riboprobe complex are added...stain the cells that contain this mRNA-riboprobe-antibody complex and, as a result, these cells turn blue...
  17. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    Published
    Aug. 30, 2016, 2:30 p.m.
    ... RNA amplification, new toehold switch designs, CRISPR/Cas9-mediated genotyping, as well as our electronic... synthetic gene networks for the detection of antibiotic resistance and strain-specific diagnostics for...electronic reader that provides portable quantification of diagnostic results.  Our approach now detects samples...
  18. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    Published
    Oct. 27, 2015, 3 p.m.
    ... genes for proteins in these compartments using CRISPR systems should strongly consider using moxFPs over...by guest bloggers Erik L. Snapp and Lindsey M. Costantini. "You underestimate the power of the Dark Side...non-fluorescent, and misfolded FP-fusions on quantitative imaging experiments and cell physiology in general...population is not readily apparent and can confound quantitative imaging experiments or even negatively impact...misfolded dark FP fusions including erroneous quantitation of fusion protein levels. It remains unclear...proteins. Our recent publication illustrated the quantitative increase in fluorescent signal achieved when...our guest bloggers Erik L. Snapp and Lindsey M. Costantini! Erik Lee Snapp received his PhD in Molecular...
  19. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    Published
    June 14, 2018, 11:44 a.m.
    ...- a primer for beginners Adenoviral delivery of CRISPR/Cas9 5 tips for troubleshooting viral transductions...formation assay on A549 cells . Alternatively, antibody-based assays are commercially available for faster...ratio for the purified virus should be ~1.3 Quantitative PCR: the number of viral DNA packaged in virions...virions is determined by a standard curve of known quantity and primers specific for a viral DNA sequence....
  20. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    Published
    Oct. 23, 2018, 11:49 a.m.
    ...packaging transfections Find tips for a first time CRISPR user Resources on Addgene.org Learn how Addgene...titration by qPCR Overview: AAV titering by qPCR quantifies the number of genome-containing viral particles... primer/probe set can be used and samples are quantified by comparing to an AAV standard curve, which ...Universal real-time PCR for the detection and quantification of adeno-associated virus serotype 2-derived...
Showing: 161 - 180 of 197 results