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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 243 results for: GRN

Showing: 101 - 150 of 243 results

  1. 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... 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...

  2. 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 ...A in any of the reading frames downstream of the gRNA target site so that transcripts undergo nonstop ...

  3. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    ...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 ...

  4. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    ... 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:...

  5. 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...gene contained within your insert (or targeted by a gRNA/shRNA) and select the species that the gene or insert...

  6. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    ...target DNA sequence as directed by the guide RNA (gRNA), while the CRY2PHR-activator fusion floats freely...occurs. Bubeck et al. expressed a telomere-targeting gRNA and a dCas9 fusion protein with three red fluorescent...Photocleavable ssDNA oligonucleotides Cas9, photocaged-gRNA Gene editing Manna et al., 2019   NPPOC-caged ..., 2020   NPOM-caged nucleobases Cas9, photocaged-gRNA Gene editing For more details, check out this...

  7. Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]

    Type
    Blog Post
    ...activity coupled with a customizable single guide RNA (sgRNA) that bears complementarity to a 20bp target sequence...multidirectional control, a new study repurposes the sgRNA to include modular synthetic RNA scaffolds that ...modules are comparatively small allowing multiple sgRNA constructs to be introduced simultaneously in a ...

  8. Plasmids 101: NGS Quality Control for Pooled Libraries

    Type
    Blog Post
    ...re-amplified our distribution stock of the Brunello Human gRNA library, and we thought it would be a good time ... the plasmids are all very similar sizes, like a gRNA library, this is less of a concern, but Addgene ...confirmed using next-generation sequencing (NGS). For gRNA libraries, you’ll design primers to create 200-300...

  9. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    ... a look at All_in_one_CRISPR/Cas9_LacZ, a CRISPR gRNA plasmid from Lynne Postovit’s lab. It contains an...it won’t tell you whether the vector contains the gRNA insert or not. A blue-white screen, however, can...and distinguish between plasmids that contain the gRNA (white) from those that do not (blue). Let’s take...

  10. Choosing a CRISPR Nuclease: Site Accessibility, Specificity, and Sensitivity

    Type
    Blog Post
    ...site that is not a perfect match to the guide RNA (gRNA) used to target the genomic locus of interest. As...of Sniper-Cas9 to different Cas9 variants using gRNAs of different lengths (Lee et al., 2018) and found...and sensitivity across a large number of loci and gRNA lengths. Recent alternatives to Cas9 While Cas9 ...

  11. Hot Plasmids - October 2020

    Type
    Blog Post
    ... Cas9-Cas12a gRNAs under a single U6 promoter. The paralog and dual-targeting hybrid gRNA library can ...interactions between paralogs. The exon-deletion hybrid gRNA library deletes human alternative cassette exons...

  12. Hot Plasmids February 2024

    Type
    Blog Post
    ...endosomal escape. Combining Cas9-PAGE with lentiviral sgRNA expression yielded editing of 80–90% in human cell...limited (likely due to its long, negatively charged sgRNA), RNP delivery was much better with Cas12, which...Workflow to assess Cas9-PAGE editing with viral sgRNA targeting mCherry reporter (mChe), with co-incubation...

  13. 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...promoters, antibiotic resistance selection markers, gRNAs, and dCas9 proteins. Second, it can be introduced...

  14. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog Post
    ...Then choose which Cas enzyme to use and design a gRNA to where you would like to introduce the edit. Finally... (Yu et al., Nat Chem Biol). Mutate the PAM or sgRNA sequences Cas9 likely goes through multiple rounds....org Browse All CRISPR Plasmids Find Validated gRNAs for Your Next Experiment Check out our CRISPR Guide...

  15. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ...multiplexing; expressing 5 gRNAs simultaneously produces comparable gene knockdown to single gRNA expression. Although....org Browse All CRISPR Plasmids Find Validated gRNAs for Your Next Experiment Check Out Our CRISPR Guide...

  16. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ... E and the N2 gene of SARS-CoV-2. If the Cas12a-sgRNA complex binds to the target sequences, Cas12a activity... Cas13, upon finding complementarity between the gRNA and a given fluorescently labelled substrate, gets...Fluorescently labelled dCas9 complexes containing sgRNA that have no mismatches bind to the immobilised ...

  17. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ...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...Addgene.org Browse All CRISPR Plasmids Find Validated gRNAs for Your Next Experiment Check out our CRISPR Guide...

  18. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ...many software tools predict off-target effects of sgRNAs using computational methods. They identify possible...mismatches based on the sequences of the genome and sgRNA. This is a good starting point for most researchers...position of the mismatch along the length of the gRNA given the stricter sequence requirement at the terminal...

  19. The Twelve Days of CRISPR

    Type
    Blog Post
    ...Addgene gave to me: tips and tricks for designing gRNAs for CRISPR editing. On the fourth day of CRISPR,... day of CRISPR, Addgene gave to me: multiplexed gRNAs.  We hope you’ve been enjoying our CRISPR blog series...

  23. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    ...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...

  24. CASTing Off for New Shores in Human Genome Editing

    Type
    Blog Post
    ...system identifies and binds a genomic target using a gRNA, and an affiliated TniQ protein recruits transposition...integrate large sequences into genomic DNA downstream of gRNA-specified sites. Created with BioRender.com. ...

  25. 10 Great Guest Posts We're Thankful For

    Type
    Blog Post
    ...use CombiGEM to barcode small libraries of CRISPR gRNAs to use for screening experiments. Get the full details...modular RNA binding proteins, dCas9, and a modified gRNA to direct a variety of customizable functions to...

  27. Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

    Type
    Blog Post
    ... plasmid or in vitro transcribed, and changes in gRNA secondary structure can affect editing efficiency...will represent a key advantage over Cas9, for which gRNA sequence can greatly affect editing efficiency. ...

  28. Another Pathway into Cells: iTOP

    Type
    Blog Post
    ...iTOP efficiently took up both Cas9 protein and a gRNA (yes, iTOP can also be used for RNA transduction...of iTOP. iTOP can be used to transduce Cas9 and a gRNA, which subsequently enter the nucleus and cleave...

  30. Targeted Mutagenesis with EvolvR

    Type
    Blog Post
    ...for targeted mutagenesis of a site of interest. A gRNA is used to direct the PolI3M-nCas9 complex to a ...window also increased the likelihood that a single gRNA would introduce more than one mutation near the ...

  31. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    ...shRNA sequence specific to your gene’s mRNA or a sgRNA targeting an exon in your gene of interest, for ...with many applications. These libraries include sgRNAs targeting the entire genome, overexpression of ...

  32. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...your protein of interest. CRISPR-Cas9 and a custom gRNA is all you need to generate a frameshift mutation... to a dead Cas9. The system is led, via a custom gRNA, to your gene of interest to temporarily increase...

  34. Plasmids 101: Modular Cloning

    Type
    Blog Post
    ...many gRNAs delivered with a relevant Cas nuclease. MoClo can be used to assemble multiplexed gRNA systems...

  37. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    ... system uses light to deactivate photocleavable gRNAs within seconds. A CRISPR-based lineage tracing tool...cerevisiae genes with 6-12 guides per gene where gRNA expression is induced by anhydrous tetracycline....

  38. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    ...those generated by multiplexed delivery of multiple sgRNAs. Since multiplexing is commonly used to create ...helps the researcher see which of the multiplexed sgRNA was involved in a particular edit, and which type...

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

    Type
    Blog Post
    ...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...

  44. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    ...nickase) fused to a deaminase that makes the edit A gRNA targeting Cas to a specific locus A target base ...CRISPR plasmids for base editing Find validated gRNAs ...

  45. The 12 Days of CRISPR: 2021

    Type
    Blog Post
    ... day of CRISPR, Addgene gave to me: engineered pegRNAs to improve prime editing efficiency. On the third...

  49. Editor's Choice, August 2016

    Type
    Blog Post
    ...the transitions at target sequences specified by gRNAs. Caveats, potential specificity issues, and other...
Showing: 101 - 150 of 243 results