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We narrowed to 177 results for: grna design

Showing: 61 - 80 of 177 results

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

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    Blog Post
    ...2.0, with two gRNAs targeting each site. They found that DNA sequences targeted by the gRNAs were the top...enrichment of DNA targeted by a negative control gRNA or predicted gRNA off-target sites. Long-range chromatin ... for Your Next Experiment Learn More About Designing gRNAs Against Your Favorite CREs Here Additional...added biotin acceptor site BirA, a biotin ligase a gRNA(s) targeting a single genomic location of interest...When these three components are co-expressed, the gRNA targets the dCas9 to the loci of interest and dCas9...cell lines will have varying levels of dCas9 and gRNA expression. Additionally, CAPTURE 3C-based sequencing... Lentiviral delivery of BioTAP-tagged dCas9 and gRNAs. Using lentiviral delivery of the two remaining ...

  2. CRISPR Plasmids - Drosophila

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    Collection
    ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR plasmids have been designed for use ...increase transcription of a specific gene. Design your gRNA sequence to direct the dCas9-activator to ...choose does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...Insert Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors...-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas protein...CRISPR plasmids that have been designed for use in Drosophila and other insects. CRISPR...

  3. CRISPR Plasmids - Zebrafish

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    Collection
    ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR plasmids have been designed for use ...Type Promoter PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors...-specific gRNA in the same cell at the same time. Single plasmids containing both the gRNA and Cas protein...CRISPR plasmids that have been designed for use in zebrafish. CRISPR...double-strand break (DSB) at a specific location based on a gRNA-defined target sequence. DSBs are preferentially...flanking the DSB and a specific edit close to the gRNA PAM site. When a repair template is present, the...

  4. CRISPR Plasmids - Plants

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    Collection
    ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR plasmids have been designed for use ...increase transcription of a specific gene. Design your gRNA sequence to direct the dCas9-activator to ...choose does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...expression by interfering with transcription. Design your gRNA to target your gene of interest’s promoter...using does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...Selectable Marker PI Publication Empty gRNA Expression Vectors Select a gRNA expression plasmid based on factors...

  5. Cpf1 Update: Comparison to Cas9 and NgAgo

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    Blog Post
    ...examine Cpf1 off-target editing. First, they designed gRNAs with single and double mismatches throughout...tolerate mismatches at gRNA positions 1, 8, 9, and 19-23. Accordingly, the 3’ end of the gRNA target sequence...kb, SpCas9 barely fits into AAV when you add in a gRNA and regulatory elements. To solve this problem, ...well into AAV vectors, especially given that Cpf1 gRNAs are also shorter than those needed for Cas9 (42 ...: Effects of (A) double and (B) single base pair gRNA-target mismatches on modification by AsCpf1 (red...Cas9 orthologs, Cpf1 efficiency varies widely with gRNA sequence. Both groups used multiple methods to examine...not mediate detectable off-target cleavage. Most gRNAs directed low-frequency Cpf1 cleavage at 1-12 off-target...

  6. Mapping the 4D nucleome with CRISPR/Cas9

    Type
    Blog Post
    ... PAM sequences required by each ortholog, a gRNA designed for one dCas9 should be specific to that ortholog...adapted to fluorescently label a sequence matching its gRNA, facilitating the live study of chromatin dynamics...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...

  7. Hot Plasmids and Viral Preps - July 2021

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    Blog Post
    ...CRISPR inhibition (CRISPRi). A) Lentiviral vector design. B) Strategy for leak-free transcriptional control...Organoid EasyTag workflow combines thoughtfully designed plasmid components with Fluorescence Activated... heavy optimization. Claudia Vickers’s lab has designed novel SL fluorescent biosensors by integrating...biosensors could have a major economic impact.   Design and mechanism of action of the receptor-based SL...Kimmelman targets mouse metabolic genes with ~6 gRNAs per gene.  CRISPR-Act3.0, developed by Yiping Qi's...Shirley Liu's lab targets all mouse genes with 10 gRNAs per gene. The guides are optimized to maximize on-target...

  8. Hot Plasmids - August 2020

    Type
    Blog Post
    ...RNA (gRNA) sequences. But these approaches rely on continual expression of both dCas9 and the gRNA. This...track cells expressing both dCas9 and gRNAs. The multi-gRNA piggyBac vector integrates into genomic...expression can vary significantly depending on transgene design and delivery. To overcome these variations in human...and sustained expression of single or multiplexed gRNAs and of dCas9 fusions. The dCas-KRAB activator and... driven by a doxycycline inducible promoter. The gRNA piggyBac plasmids include a mRFP reporter. With ...protein levels. Plasmids for multiplexed piggyBac gRNA delivery and the all-in-one dCas9-KRAB and dCas9...at Addgene. Check out these plasmids for piggyBac gRNA delivery and the all-in-one dCas9 targeting plasmids...

  9. CRISPR Plasmids - Parasites

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    Collection
    ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Eukaryotic parasites ...Toxoplasma gondii CRISPR genome-wide knockout gRNA pooled library designed with 10 guides against 8,158 predicted...CRISPR expression plasmids that have been designed for use in parasites. CRISPR...double-strand break (DSB) at a specific location based on a gRNA-defined target sequence. DSBs are preferentially...flanking the DSB and a specific edit close to the gRNA PAM site. When a repair template is present, the...

  10. CRISPR Plasmids - Prime Edit

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    Collection
    ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...prime editing guide RNA (pegRNA). As with a typical gRNA, the pegRNA is designed with a spacer that binds...find a gRNA vector based on expression system, promoter, the type of gRNA (e.g., pegRNA, epegRNA, nicking...post . Need help designing a pegRNA? Try the DeepPrime (Link opens in a new window) design tool from Hyongbum...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Prime editing is a “search...Publication Empty Prime Editing gRNA Vectors A selection of empty gRNA vectors suitable for prime editing...PEmax to enhance pegRNA stability epegRNA - additional protection added to 3’ tail of pegRNA to prevent RNA...

  11. CRISPR Plasmids for Genomic Visualization

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    Collection
    ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Using catalytically inactive...is easy to implement due to the simplicity of gRNA design, programmable for different genomic loci, capable...multiple gRNAs that tile the locus. Chromosome painting requires delivery of hundreds of gRNAs with target...fluorescence microscopy in living cells. Alternatively, gRNAs can be fused to protein-interacting RNA aptamers...efficiently visualized in living cells using a single gRNA that has multiple targeted sequences in close proximity...

  12. CRISPR Plasmids - Repress Gene Expression

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    Collection
    ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Catalytically dead dCas9...expression by interfering with transcription. Design your gRNA to target your gene of interest’s promoter...using does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...

  13. Design Tips for Prime Editing

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    Blog Post
    ...principles pegRNA design If you’re just starting out with prime editing, you’ll need to design and optimize...great tips to guide your experimental design, especially the design of the RNA sequences you’ll use in prime... a prime editing guide RNA (pegRNA). The pegRNA consists of a CRISPR gRNA extended to include a primer...degradation. When designing epegRNAs with large structured motifs like mpknot, use the pegRNA Linker Identification...extension of the pegRNA should not be C. A C base is speculated to base pair with G81 of the gRNA (Anzalone ...PE3b and PE5b: In this approach, the nicking sgRNA is designed to bind only after the edit is installed. ...PE5 approach. Figure 7: Nicking sgRNAs can be designed to target edited DNA, so that nicking only...

  14. CRISPR Plasmids - Activate Gene Expression

    Type
    Collection
    ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Catalytically dead dCas9...increase transcription of a specific gene. Design your gRNA sequence to direct the dCas9-activator to ...choose does not also express a gRNA, you will need to use a separate gRNA expression plasmid to target ...

  15. CRISPR Plasmids - Purify Genomic Loci

    Type
    Collection
    ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Identifying molecules...genomic DNA bound by the gRNA. An epitope tag(s) can be fused to dCas9 or gRNA for efficient purification...interest in vivo. For CRISPR-based purification, design your gRNA sequence to direct dCas9 to a specific locus...of any genomic sequence specified by a particular gRNA. In the enChIP (engineered DNA-binding molecule-...

  16. Optimizing Donor DNA for Enhanced CRISPR Genome Editing

    Type
    Blog Post
    ... Brush Up on Your HDR Basics Learn How to Design Your gRNA Find the Ideal CRISPR Software for Your Needs...Plasmids for Your Research Check out Our List of gRNA Design Tools ... single or double stranded donor DNA with other design parameters. Example single stranded donor DNA and...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...

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

    Type
    Blog Post
    ...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...’s what our plasmids allow.” Wang’s library is designed to target a smaller set of genes and it includes...sequential infection first of Cas9, then with the sgRNAs, and can result in a higher titer due to the smaller...

  18. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ...generate a knock-in.   General considerations for designing a repair template to create mutations HDR donors...that Cas enzymes can continue to cleave DNA if the gRNA/PAM site remains intact. In other words, Cas9 will...resulting mutations after an HDR event, ensure the gRNA or PAM site is disrupted in the donor. Choosing...increase the frequency of edits, researchers have designed self-cleaving plasmids that liberate the targeting...information on HDR and CRISPR. Our CRISPR 101 series was designed to help explain the basic principles driving CRISPRs...et al. PLoS ONE 9(8); (2014). PubMed. Making designer mutants in model organisms. Peng, Y. et al. Development...Addgene.org Browse All CRISPR Plasmids Find Validated gRNAs for Your Next Experiment Check out our CRISPR Guide...

  19. CRISPR Plasmids - Epigenetics

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    Collection
    ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts To make targeted epigenetic...catalytically dead dCas9 to epigenetic modifiers. Design your gRNA to target a specific promoter or enhancer ...

  20. CRISPR Plasmids - Tagging

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    Collection
    ... dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila...expresses Cas9 & PITCh-gRNA pX330A-FBL/PITCh - expresses Cas9, PITCh-gRNA, & FBL-specific gRNA pCRIS-PITChv2-FBL...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The CRISPR system has...containing Cas9 and a validated gRNA, based on the Zhang lab's PX458 . Multiple gRNA plasmids may be used. The...currently unavailable, you will need to design and clone in a gRNA(s) to guide the Cas9 protein to your ...Plasmids Protein Species Tag Donor Plasmid gRNA plasmid gRNA plasmid FLAG pFETCh_Donor CEBPB Human FLAG... for more information about HDR/repair template design. How to use CRISPR to tag your gene of interest...
Showing: 61 - 80 of 177 results