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We narrowed to 177 results for: grna design

Showing: 101 - 120 of 177 results

  1. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ... Encodes gRNA/shRNA A plasmid with one or more gRNA or shRNA sequences cloned into it. gRNAs are used...the shRNA sequence. Empty backbone A plasmid designed for cloning a gene/insert into it. Empty backbones...gene contained within your insert (or targeted by a gRNA/shRNA) and select the species that the gene or insert...intended to be used. For instance, if your plasmid is designed to allow you to produce large amounts of your ...

  2. CRISPR Plasmids - RNA Targeting

    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 Type VI CRISPR systems...

  3. Twelve Amazing Guest Blog Contributors!

    Type
    Blog Post
    ... An Introduction to Gating in Flow Cytometry, Designing Your First Flow Panel, and Beyond Surface Labeling...software that predicts efficiencies of prime editing gRNAs.   PRIDICT Harvinder Virk and Michael Biddle, ...

  4. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    Blog Post
    ...single-target design”, “multi-target design”, “off-target aware”, “high mismatch limit”, “approximate design”, “... “empirical design”, “single-PAM design”, and “multi-PAM design”. Trends: By the end of 2014, tools began...with the post-design process such as primer and plasmid design. Examples:  “multiplex design”, “multi-method...what determines good sgRNA design. Incidentally, the first software for sgRNA design (by Hsu et al., 2013...results to the designs suggested by each tool. For those who are entering sgRNA design for the first time...multi-method design”, and “single-method design”. Trends: The most common utility functions are batch design...aiding in primer design are more and more common. User interaction: Software design elements that fall...

  5. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ...treated with a folA gRNA were more sensitive to trimethoprim when compared to control gRNA treated or untreated...existing bacterial CRISPRi methods. First, its modular design allows for customization with strain-specific promoters...promoters, antibiotic resistance selection markers, gRNAs, and dCas9 proteins. Second, it can be introduced...

  6. CRISPR Plasmids - CRISPR Transposases (CAST)

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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 CRISPR-associated transposases...

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

    Type
    Blog Post
    ... 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 ...HDR event can’t occur. Lastly, depending on its design, donor A may be capable of expressing a truncated...A in any of the reading frames downstream of the gRNA target site so that transcripts undergo nonstop ...

  9. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    ...strategies for Duchenne muscular dystrophy. gRNAs can be designed to flank a single or multiple exons that...In a subsequent study, they used CRISPR with two gRNAs to delete exon 51 or exons 45-55 in patient myoblasts...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 ...

  10. Pushing the Limits of DNA Assembly

    Type
    Blog Post
    ...multiple fragments of DNA to create a synthetically designed DNA sequence. There are multiple methods of DNA...purification steps and end-repair steps with well-designed fragment overlap sections, while still creating... guide you through both experimental and primer design based on fragment sequences and the polymerase ...enzymes are adjusted. Appropriate fragment and primer design results in covalently joined, scarless DNA – perfect...interest (search Addgene plasmids by gene here), designing primers with your desired mutation, performing...Addgene.org Browse Our CRISPR Resources Find Validated gRNAs Browse All Plasmids ...thousands of base pairs. Cloning sgRNAs Generating constructs to produce sgRNA for CRISPR experiments can be...

  11. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    ... packaging Streptococcus pyogenes (SpCas9) and a gRNA together (~4.2 kb) into an AAV vector is challenging...Zhang's group previously packaged Cas9 and multiple gRNAs into separate AAV vectors, increasing overall packaging...regulatory sequences while still fitting Cas9 and its gRNAs into one AAV, Cas9 must be made smaller. Previous...from Streptococcus thermophilus and a rationally-designed truncated Cas9. Unfortunately, certain drawbacks... the Zhang lab created an all-in-one SaCas9 and sgRNA construct using the liver-specific serotype AAV8... a control AAV-GFP. Not only did the AAV-SaCas9-sgRNA constructs mediate genome modification, but they...

  12. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    ...at 37 °C.     The new pEcCas/pEcgRNA system We speculated that the gRNA on pCas that is specific to the...system with the updated pEcCas/pEcgRNA system, and confirmed that indeed the gRNA-pMB1 had a slightly higher...changes. In pCas9, we 1) replaced the promoter of gRNA-pMB1 with PrhaB, 2) changed the replicon of pCas...mechanisms behind their distinct phenotypes and to design a new generation of engineered cells for green ...process begins with the addition of the sgRNA sequence in pEcgRNA. Next both plasmids are transformed in...generate pEcgRNA. The ccdB serves as a counterselection marker and will be replaced by the sgRNA targeting...the newly generated pEcCas/pEcgRNA system.   Advantages of the pEcCas/pEcgRNA system Compared with the ...

  13. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ...human cells (Kim et al, 2016). The Waldman lab has designed their AAV tagging vectors (pAAV-SEPT-Acceptor ...vectors for your specific gene, the lab recommends designing homology arms with the modification of interest...use a dual vector system composed of SaCas9 and a gRNA in one AAV and a repair template in second a second...small Cas9 homologs like SaCas9 and their requisite gRNAs take up essentially all of an AAV’s ~4.5 kB packaging...

  14. Validated gRNA Sequences

    Type
    Collection
    ...Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts The table below lists gRNA sequences that have...pyogenes, S. aureus etc.) was this gRNA sequence designed for? A given gRNA sequence may only be compatible...indicate the Cas9 application the gRNA was designed to accomplish. Validated gRNA Sequence Datatable Target Gene... Genome Engineering CRISPR Validated gRNA Sequence Datatable Validated gRNA Target...Does a given gRNA sequence exactly match your genomic target? Variation between a given gRNA sequence and...details and outcomes. Submit Validated gRNA Sequences Validated gRNA sequences can be added to this table...validated gRNA sequences. Genome...

  15. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ...replacing it or disrupting it with an piece of DNA designed in the lab. Specially constructed plasmids can...interest is difficult to target specifically with a gRNA). Be sure to keep these techniques in mind when ...locus with homologous sequence using a specially designed vector that contains sequence homology to the ... the process, we’ll walk through an experiment designed to knock out exon 2 of a given gene.   ... Figure 1: A knockout targeting vector designed to insert a resistance gene. The vector contains a neomycin... to select against random recombinants.   Design your targeting construct. For recombination to ... Figure 2: A knockout targeting vector designed to insert and later remove a resistance gene. ...

  16. Plasmids 101: Modular Cloning

    Type
    Blog Post
    ...many gRNAs delivered with a relevant Cas nuclease. MoClo can be used to assemble multiplexed gRNA systems...cloning, or MoClo, is a systematic cloning strategy designed by synthetic biologists to perform large scale...

  17. CRISPR Pooled gRNA Libraries

    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 CRISPR is a useful tool...screening experiments, due to the relative ease of designing gRNAs and the ability to modify virtually any genetic...Type Species PI Lentiviral Generation gRNAs per gene Total gRNAs Adamson DNA Repair CRISPRi Libraries ...Reduced Double-gRNA Library 137999 Knockout Human Parts 3rd 3 59,576 Human Whole Genome sgRNA iBAR Library...thousands of plasmids, each containing multiple gRNAs for each target gene. In a CRISPR screening...libraries require the use of lentivirus to deliver the gRNA/Cas9 to target cells. Therefore, you must be equipped...

  18. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ...subsequently tested the original BE3 and HF-BE3 using a gRNA with high off-target cutting, and they found that...codons) with kanamycin cassettes and were carefully designed to allow for easy removal of the resistance cassette... 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...frequently and has developed a website to aid in plasmid design and construction. We look forward to the many plant... a single, unique, and constitutively expressed sgRNA targeting the 5’ end of each essential gene. Koo...

  19. Lentivirus Plasmids

    Type
    Collection
    ...plasmids to express your CRISPR gRNA? Please see our CRISPR Empty gRNA plasmids table and search "lenti...lentiviral systems available at Addgene have been designed for increased researcher safety. Read our Lentiviral...Tet-inducible lentivirus for ORF expression, multicistronic design enables tracking of viral transduction. Also see... based) for inducible knockdown, multicistronic design enables tracking of viral transduction. See article...
Showing: 101 - 120 of 177 results