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Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

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We narrowed to 182 results for: GRN

Showing: 81 - 100 of 182 results

  1. Hot Plasmids: Fall 2024

    Type
    Blog Post
    Published
    Nov. 21, 2024, 1:30 p.m.
    ...recombination methods). By using mCherry for selection of sgRNA plasmid transformants, they avoid M. abscessus’ ...Using an inducible Cas9 expression and removing the sgRNA plasmid after identifying successful knockouts limits... KanR. This strain is then transformed with the sgRNA cassette on a plasmid expressing mCherry and resistance... of the target gene. Lastly, curing the mCherry-sgRNA plasmid leaves an edited strain carrying Cas9 to...to allow the introduction of additional sgRNAs. Image reused from Neo et al., 2024 under CC-BY license...

  2. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    Published
    Aug. 18, 2016, 2:30 p.m.
    ...the gene into a gRNA finder such as WU-CRISPR (Wong et al., 2015)) or our lab’s sgRNA scorer 1.0 (Now ...uses for Cas9 activators is in genetic screening. gRNAs targeting every gene in the human genome for example...targeted by the activator by simply providing a new gRNA rather than engineering an entirely new protein....difficult to construct and deliver when compared to gRNAs. Additionaly, cDNAs cannot be used to study in cis...

  3. Introducing an All-in-One CRISPR/Cas9 Vector System for Multiplex Genome Engineering

    Type
    Blog Post
    Published
    Nov. 12, 2014, 3:54 p.m.
    ... to Design Your gRNA for CRISPR Genome Editing CRISPR-Cas9: Tips for Optimizing sgRNA Activity Lentiviral...Cas9 vectors expressing up to seven guide RNAs (gRNAs). Sakuma said that the new system could also be ...cloning method to ligate expression cassettes of the gRNAs into a single vector in tandem. They went on to ...

  4. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    Published
    Dec. 17, 2020, 1:30 p.m.
    ...Addgene's CRISPR Resources Learn about gRNA design and gRNA optimization for your CRISPR experiment ...depending on perspective – to work with inefficient sgRNAs in my initial direct screening efforts, which were...genome editing by CRISPR whether it be by improved gRNA or repair template design, delivery, screening, ...single plasmid targeting vectors that encode the gRNA transcript and the repair template of your choosing...

  5. Your Top Requested Plasmid in 2015!

    Type
    Blog Post
    Published
    Dec. 30, 2015, 3:30 p.m.
    ...transfer vector used to deliver both Cas9 and your gRNA of interest to mammalian cells. It was designed ...use lentiCRISPR v2 as the backbone for their many gRNAs. The high titer lentivirus production afforded by...pooled libraries containing thousands of pre-cloned gRNAs. DNA aliquots of the GeCKO v2 pooled libraries targeting...Screening Using CRISPR/Cas9. If you need help cloning a gRNA into lentiCRISPR v2, you can take a look at the ...

  6. What's New in CRISPR - September 2019

    Type
    Blog Post
    Published
    Sept. 10, 2019, 12:56 p.m.
    ...targeting requires two sgRNAS: one that targets the knock-in site and a universal sgRNA that targets the donor... the genomic loci of interest. As the universal sgRNA is not predicted to target sites in the human, pig...lab. The Bikard library contains ~93,000 unique gRNAs that target random positions along the E. coli MG1655...

  7. Mapping the 4D nucleome with CRISPR/Cas9

    Type
    Blog Post
    Published
    Aug. 11, 2015, 2:30 p.m.
    ...adapted to fluorescently label a sequence matching its gRNA, facilitating the live study of chromatin dynamics...in the PAM sequences required by each ortholog, a gRNA designed for one dCas9 should be specific to that...orthologs. Ma et al. tested their dCas9 variants using gRNAs specific for telomeric sequences and showed that... label two different pairs of chromosomes using gRNAs specific to sequences on chromosomes 9 and 13. They...

  8. Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening

    Type
    Blog Post
    Published
    Feb. 20, 2014, 4:57 p.m.
    ...protein coding regions with 87,897 unique gRNAs. Their gRNA library is delivered via lentiviral infection...Sabatini and Eric Lander, comprise 73,171 guide RNAs (gRNAs) covering a total of 7,114 human genes. Unlike the...sequential infection first of Cas9, then with the sgRNAs, and can result in a higher titer due to the smaller...

  9. New CRISPR Web Resources and #12DaysofCRISPR Recap

    Type
    Blog Post
    Published
    Dec. 12, 2017, 3:13 p.m.
    ...- see our recap below! Day 1: gRNA design tools: How to Design your gRNA for CRISPR Genome Editing and...Cas9 variants: CRISPR 101: Multiplex Expression of gRNAs and The PAM Requirement and Expanding CRISPR Beyond...HypaCas9 Day 10: Transcriptional regulation: Truncated gRNAs for Regulating Gene Expression and CRISPR 101: Epigenetics...

  10. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    Published
    Sept. 21, 2021, 1:15 p.m.
    ...Veening is now available with one sgRNA per operon or gene. The sgRNA pool can also target core operons... cell lines. The Turner Lab Human messenger-RBP sgRNA Library targets human mRNA binding proteins with...operons in other pneumococcal strains. The BARBEKO sgRNA library uses CRISPR cytosine base editors to disrupt...

  11. Optimizing Donor DNA for Enhanced CRISPR Genome Editing

    Type
    Blog Post
    Published
    March 24, 2016, 2:30 p.m.
    ...research with a simple question: how do Cas9 and sgRNA interact with and dissociate from a DNA target? ..., requiring expressed Cas9 protein, transcribed sgRNA, and optional donor DNA. Chris Richardson is a...Brush Up on Your HDR Basics Learn How to Design Your gRNA Find the Ideal CRISPR Software for Your Needs Resources...Plasmids for Your Research Check out Our List of gRNA Design Tools ...

  12. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    Blog Post
    Published
    Jan. 25, 2024, 2:15 p.m.
    ...properties (CasID): Cis/trans activity, PAM requirement, gRNA, and multiplex ability Experimental guidelines ...Target requirements As we all know, a Cas guided by a gRNA is useless without an adjacent PAM or protospacer-flanking...etc.) as well as their orientation relative to the gRNA (5’ or 3’). In some cases, there is no requirement... your search (trans activity, multiplex ability, gRNA design, etc.). With this feature, you can compare...

  13. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    Published
    Feb. 11, 2025, 2:15 p.m.
    ...Cascade is brought to the target location using a gRNA and can then recruit Cas3, which carries out the... To get large deletions using Cas9, two or more gRNAs must be used along the same chromosome. During DNA...will repair without the section between the two gRNAs, creating a deletion. However, there is the chance... the targeted portion of DNA using only a single gRNA. Cas3 appears to have fewer off-target alterations...

  14. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    Published
    March 28, 2018, 6:52 p.m.
    ... PAM near your insertion site, the corresponding gRNA might be undesirable due to potential off-target...activity. Surprisingly, off-target effects across 5 gRNAs with NGG PAM sequences were actually lower for xCas9...3.6/3.7 than for SpCas9. For some non-promiscuous gRNAs, xCas9 3.7 displayed more than a 100 fold decrease...decrease in off-target editing; for known promiscuous gRNAs, xCas9 3.7 still showed a 4.2-9.4-fold improvement...

  15. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    Published
    Nov. 29, 2022, 2:15 p.m.
    ...indicating loci where SNPs reside on sgRNA target sequences for sgRNAs that target ancestry-associated genes...that could analyze CRISPR sgRNAs and report the likelihood of any given sgRNA having an ancestral bias....

  16. 15 Hot Plasmids from 2017

    Type
    Blog Post
    Published
    Jan. 9, 2018, 3:02 p.m.
    ...subsequently tested the original BE3 and HF-BE3 using a gRNA with high off-target cutting, and they found that... a single, unique, and constitutively expressed sgRNA targeting the 5’ end of each essential gene. Koo... constructs (wild-type and truncation mutants), gRNAs used for TZAP gene editing, and a Hi6xs-MBP tagged...as Cys4, tRNA, and ribozyme systems for multiplex gRNA expression. Expression of toolkit components can...compatible backbones that can be combined with your gRNA and repair templates of choice though modular cloning...

  17. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    Published
    Sept. 1, 2020, 1:15 p.m.
    ... for integration, “part A” of the transgene, the gRNA target site that mediated the donor template’s integration...integration; and a sequence of stuffer DNA after the gRNA target site. Donor B contains: 400 bp homology ...if non-homologous end joining (NHEJ) disrupts the gRNA target site after the first HDR event, then the ...A in any of the reading frames downstream of the gRNA target site so that transcripts undergo nonstop ...

  18. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    Published
    Jan. 26, 2016, 3:30 p.m.
    ...In a subsequent study, they used CRISPR with two gRNAs to delete exon 51 or exons 45-55 in patient myoblasts...skipping strategies for Duchenne muscular dystrophy. gRNAs can be designed to flank a single or multiple exons...NHEJ, will restore the dystrophin reading frame. gRNAs bind in intronic regions, so low-frequency indels...CRISPR Plasmids for Your Research Find Validated gRNAs for Your Research ...

  19. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    Published
    March 25, 2025, 1:15 p.m.
    ... Acronym Full name Notes CRISPR terms gRNA/sgRNA Guide RNA RNA used by Cas9 to locate a matching... the DNA strand that does not base pair with the gRNA. Base editors target this “non-target strand.” To... predicting CGBE editing outcomes and assisting sgRNA selection is available at http://www.sunlab.fun:...

  20. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    ... Encodes gRNA/shRNA A plasmid with one or more gRNA or shRNA sequences cloned into it. gRNAs are used...gene contained within your insert (or targeted by a gRNA/shRNA) and select the species that the gene or insert...
Showing: 81 - 100 of 182 results