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We narrowed to 108 results for: cas9 variant

Showing: 61 - 80 of 108 results

  1. CRISPR Challenges: Standardization and Homology Directed Repair

    Type
    Blog Post
    ... Check out this post to see if an engineered Cas9 variant might work for your experiment. PITChing MMEJ...Corn’s lab describe how a better understanding of Cas9 biophysics can help enhance HDR. CRISPR 101: Ribonucleoprotein...tool to help you design, pick, and close guides for Cas9 and Cpf1. This tool supports over 250 genomes, and...The PAM Requirement and Expanding CRISPR Beyond SpCas9 The closer your edit is to the PAM site, the higher...

  2. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    ... a colorful Cas9 toolbox, Ma et al. turned to SpCas9 and its orthologs NmCas9 and St1Cas9. Each ortholog...locus, and to do so in vivo. Catalytically dead Cas9 (dCas9) cannot induce gene editing, but it retains its...the other orthologs. Ma et al. tested their dCas9 variants using gRNAs specific for telomeric sequences...success of Ma et al.’s approach, fluorescently labeled Cas9’s come with a number of limitations. Since color...by PAM sequence, each color requires a different Cas9 ortholog, and the target sequence must be located...genome editing. If used with catalytically active Cas9, the short, CRISPRainbow gRNA will permit labeling...publication at Addgene. 4. Kiani, Samira, et al. “Cas9 gRNA engineering for genome editing, activation ...

  3. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...pDGE Dicot Genome Editing Kit contains Cas9, Cas9 nickase, and Cas9 activator expressing plasmids that can...CRISPR-Cas9-based tool for targeting CpG methylation by fusing catalytically inactive Cas9 (dCas9) with...high-throughput CRISPR/Cas9 gene modification in Caenorhabditis elegans CRISPR/Cas9 technology has revolutionized...reported on a novel Cas9-based activator developed by the Church lab, termed “dCas9-VPR”, which facilitates...gRNA expressing cells were then transfected with a Cas9-expressing plasmid to generate a population of mutant...post entitled “Genome-wide Screening Using CRISPR/Cas9”. Detailed information regarding the Wu Lab library...gene was chosen to represent the primary splice variant in 38 types of human cancer. Many of the transcripts...

  4. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

    Type
    Blog Post
    ...Screening immune cell lineages in vivo using the CRISPR-Cas9 system CHIME Article contributed by Aliyah Weinstein...custom nucleases for each gene of interest. CRISPR-Cas9 tools have been used in immunology for in vitro ...Medical School developed a solution: CHIME, a CRISPR-Cas9 system that can edit genes in any immune lineage...in vivo, the non-immunogenic violet-excited GFP variant Vex. To understand the impact of this gene editing...expressed this construct in bone marrow stem cells from Cas9-expressing mice, and used the cells to reconstitute...

  5. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

    Type
    Blog Post
    ...the Addgene Blog Learn about High Fidelity Cas9 Variants How to Design Your gRNA Delivering CRISPR Ribonucleoproteins...at the Rockefeller University in NYC. The CRISPR/Cas9 system is a versatile tool for precise gene editing...organisms and model systems. We have used CRISPR/Cas9 extensively for the purpose of making sequence-specific...induced pluripotent stem cells (iPSCs). The CRISPR/Cas9 com­plex is very efficient at introducing double...Cas-blocking mutations After genome editing with CRISPR/Cas9 in human cells was described in 2013 (Cong et al...amounts of single cell clones. Furthermore, CRISPR/Cas9 editing is often bi-allelic, meaning that even if...systematically analyzed: introducing silent CRISPR/Cas9 blocking mutations in either the PAM (changing one...

  6. New CRISPR Web Resources and #12DaysofCRISPR Recap

    Type
    Blog Post
    ...CRISPR Antimicrobials Day 7: Multiplexing and Cas9 variants: CRISPR 101: Multiplex Expression of gRNAs and...Pooled libraries: Genome-Wide Screening Using CRISPR/Cas9 and Plasmids 101: NGS Quality Control for Pooled... CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9 Day 10: Transcriptional regulation:...The PAM Requirement and Expanding CRISPR Beyond SpCas9 Day 8: Repair mechanisms: CRISPR 101: Non-homologous...

  7. Hot Plasmids - November 2023

    Type
    Blog Post
    ...such as long or highly structured templates. Cas9 variants PE6e–g often had even higher editing efficiencies...vivo. Incorporating reverse transcriptase (RT) variants, PE6a and PE6b are 500–800 bp smaller than previous...PE6d are optimized for dual-AAV systems. These variants use different RT enzymes and performed large edits...edit.   Figure 1: Choosing the best PE6 variant for your application. Image reused form Doman and...

  8. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

    Type
    Blog Post
    ... activation CRISPR activation uses dCas9, a CRISPR protein variant lacking its endonuclease ability, to...J, Collins JJ, Church GM (2015) Highly efficient Cas9-mediated transcriptional programming. Nat Methods...Perrimon N, Collins JJ, Church G (2016) Comparison of Cas9 activators in multiple species. Nat Methods 13:563...transcriptional activation by an engineered CRISPR-Cas9 complex. Nature 517:583–588 . https://doi.org/10.1038...Gersbach CA (2013) RNA-guided gene activation by CRISPR-Cas9–based transcription factors. Nat Methods 10:973–... cell lines.   dCas9-VP64 Description CRISPR activation can occur through fusing dCas9 with VP64, a strong... for CRISPR-based gene activation. dCas9-CBP In this method, dCas9 is fused to CBP, an histone acetyltransferase...

  9. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    ...option is to use variants of Cas9, like split Cas9, or a smaller Cas9 ortholog like SaCas9. You could even...at double-stranded break points caused by CRISPR-Cas9 editing. The good news is that this integration ...approximately 4.7 kb — which means that when using Cas9, you’re already almost out of space! Don’t let this... you have one AAV vector dedicated to expressing Cas9, while the other houses the gRNA and scaffold. If...even venture away from the comfort of Cas9, and opt for a completely different Cas effector, like Cas12a...

  10. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    ...time CRISPR users Learn how to Choose the Best Cas9 Variant for Your Next Experiment Learn More About Designing...Additional Resources on Addgene.org Find Other Cas9 Variants for Purifying Genomic Regions of Interest Here...co-expressed, the gRNA targets the dCas9 to the loci of interest and dCas9 is biotinylated by BirA. From here...To increase the resolution, the Xu lab created a dCas9-based CAPTURE (CRISPR Affinity Purification in situ... cell line that co-expresses three components: dCas9 with an added biotin acceptor site BirA, a biotin...be pulled down via streptavidin IP. Biotinylated dCas9 goes along for the ride throughout all of these ... CAPTURE cell lines will have varying levels of dCas9 and gRNA expression. Additionally, CAPTURE 3C-based...

  11. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ... eSpCas9(1.1), or HypaCas9. Read more about Cas9 options in our blog post here. Dealing with low editing...version of Cas9 that has been specifically engineered for reduced off target editing, such as SpCas9-HF1, eSpCas9...gRNA and introduced it into your target cells with Cas9. Hooray! Now it’s time to make sure your genome ... product of both an unedited population and your Cas9-targeted cells. Simply upload both trace files to...with the estimated editing frequency of your sgRNA-Cas9 combo. Pro tip: When designing your PCR, ensure ...introduced a donor template along with your gRNA and Cas9 combo and are hoping for homology-directed repair...despair, you have options! If you think delivery of Cas9 or gRNA is the issue, check out or blog on optimizing...

  12. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...CRISPR-Cas9 genome editing may be the hot new way to manipulate gene expression, but other gene manipulation...expression; these deletions are defined using 2 loxP variants (only identical loxP sites can mediate recombination...either deletion removes one of the other loxP site variants, preventing further recombination. Brainbow-1.1...with recombination events specified by three loxP variants, but the principles remain the same. The first...Variations Between Brainbow Systems   Brainbow variants and applications beyond the mouse brain In addition...Brainbow to be used in a variety of species. More variants have been created by other labs, including R26R-Confetti...

  13. Plant Plasmids and Resources

    Type
    Collection
    ...of plasmids expressing different functional variants of Cas9 and empty gRNA backbones. A few examples of... an intronized Cas9 series of vectors and DNA modules for assembly of CRISPR/Cas9 constructs - Sylvestre...CRISPR/Cas9 assembly for fast generation of multiple Arabidopsis mutants - Niko Geldner. CRISPR-Cas9 vector...truncatula , wheat, and barley - Daniel Voytas. A CRISPR/Cas9 Toolkit for use in a variety of plant species (e.g...or more gRNA expression cassettes - Qi-Jun Chen. Cas9 and gRNA backbone for use in rice. Includes a protocol...species and genotypes - Zhenghe Li. A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex...MoClo-compatible zCas9i Cloning Kit includes a high efficiency intron-optimized SpCas9-coding gene, zCas9i, and an...

  14. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ...Use Cas9 Activators Perform Genome-Wide Screens with CRISPR Generate Mouse Models with CRISPR/Cas9 Additional...techniques, please see “Methods for Optimizing CRISPR-Cas9 Genome Editing Specificity” by Tycko et al. 2016...improving the genome-wide specificities of CRISPR–Cas9 nucleases”
by Tsai and Joung 2016. Many thanks ...high-throughput screening strategy for detecting CRISPR-Cas9 induced mutations using next-generation sequencing...sequencing: a comparison of methodologies for KRAS variant detection." Biotechniques 53.3 (2012): 182-188....of off-target DNA cleavage reveals RNA-programmed Cas9 nuclease specificity." Nature biotechnology 31.9.... "Digenome-seq: genome-wide profiling of CRISPR-Cas9 off-target effects in human cells." Nature methods...

  15. Visualizing Genomic Loci with CRISPR-Sirius

    Type
    Blog Post
    ...fluorescent protein to various Cas9 variants, SpCas9, NmCas9, and St1Cas9. These fusions localized to specific...The catalytically dead Cas9 protein (dCas9) is well known for its ability to bind DNA targets without...inhibition, among others. Over the past few years, dCas9 has also become a robust visualization tool to study...University of Massachusetts Medical School developed a dCas9-based multicolor labeling system to visualize chromosomal... al. “Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow.” Nature...

  16. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    ... years, scientists have utilized CRISPR-Cas9 protein variants, Cas13, and Cas12a, to develop simple, portable...Cas12a (Cpf1) is a CRISPR Cas variant that can also cleave dsDNA similarly to Cas9 (Zetsche et al., 2015 )....

  17. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

    Type
    Blog Post
    ...for oxidizing environments to photoconvertible variants, it seems like GFP is always learning new things...proteins with a modular structure, constructing split variants should be relatively straightforward, but it is...versions of non-modular proteins like Cre. Could Cas9-DOG be developed soon to allow genome modification...

  18. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...plasmids available! Interested in purifying Cas9? Check out pET-28b-Cas9-His from Alex Schier's lab, designed...the lab of Yonglong Chen, pCS2-3xFLAG-NLS-SpCas9-NLS is a Cas9 expression plasmid that was used by Chen...colleagues, FokI-dCas9 expresses Fok1 nuclease domain fused to catalytically inactive Cas9 DNA-binding domain...which is 45 nm more red-shifted compared to other variants. The group further optimized Chrimson through ...off-kinetics (15.8 ± 0.4ms from 21.4 ± 1.1 ms); this new variant was named ChrimsonR. Plasmids containing Chronos...lentiviral and AAV expression vectors for certain variants. Klapoetke et al., Nat Methods 2014 Mar 11;(...gene/region of interest is a key step for CRISPR/Cas9 gene editing. Dr. Masahito Ikawa has created a GFP...

  19. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...Gram-positive bacteria (Kellenberger et al., 2013). Other variants of Spinach include Baby Spinach, a miniaturized...cellular RNA to specific intracellular locations using Cas9 and track the localization of the complex. Broccoli...short non-coding RNAs (Autour et al., 2018). Three variants of Mango, coined Mango II, III, and IV, were then...high binding affinity to TO1-biotin. Two of these variants exhibit increased brightness compared to Mango...

  20. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    ...editors use either catalytically dead Cas9 (dCas9) or a Cas9 nickase (nCas9) to avoid introducing double-strand...need from Cas9 are sequence specificity and the R-loop structure. In order to bind DNA, Cas9 requires ...expanded CGBE target scope by creating BE variants with other Cas9s that recognize different PAMs. Their computational...non-target strand of DNA for Cas9 to bind.   R-loop The structure formed by Cas9 binding to DNA and separating...PAM) on the opposite strand. Once it finds a PAM, Cas9 unwinds the adjacent DNA to create a region of separate...single-stranded DNA (ssDNA) region, using enzymes fused to Cas9 to alter the exposed ssDNA bases. Thus, base editors...window restricted to the stretch of ssDNA created by Cas9 binding. Pro tip! In CRISPR, the non-target strand...
Showing: 61 - 80 of 108 results