...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... plating, screening and generation of sgRNA. Generation of sgRNA libraries, with maximum complexity, is...constructs with multiple point mutations Cloning sgRNAs Production of Yeast Expression Cassettes Generating...synthetic oligo to generate a single-guide RNA (sgRNA). Using only one single-stranded oligo, instead ...to create a double-stranded oligo. Generating an sgRNA construct becomes as simple as combining your favorite...

...commonly used for Golden Gate cloning or for CRISPR gRNA cloning. With this more functional display, you...

... 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:...

...Resources at Addgene Get Tips on How to Design a gRNA Read Our CRISPR Guide to Catch up on All the CRISPR...Read Other Blog Posts about CRISPR Find validated gRNAs ...of what determines good sgRNA design. Incidentally, the first software for sgRNA design (by Hsu et al.,...technical expertise, thoughtful single guide RNA (sgRNA) design, and the abnormally low GC content of the...without computational support. Indeed, rational sgRNA design is not possible without relying on some kind...end of 2014, I began developing software to make sgRNA design accessible to all. At the time I thought ...  Breaking down the barriers: My view on current sgRNA design tools The goal of this post is to provide...

...Through molecular biology teamwork. The guide RNA (gRNA) specifies the editing target site on the DNA, the...

...with CasRx and either an unprocessed or processed gRNA array (22 nt spacer with 30 nt direct repeat) mediating...

...for Your Next Experiment Find Our List of CRISPR gRNA Design Tools ...

...good reason. It is guided by an RNA molecule, the gRNA, to a complementary site in the genome where it ...

... the Zhang lab expressing S. pyogenes Cas9 and a gRNA cassette. It can be used to produce a ~10-fold higher...

... system developed by Taekjip Ha’s lab where Cas9-gRNA complex binds the target DNA without cleavage until...

...CRISPR plasmids for protein tagging Find validated gRNAs ...

...variable portion of the plasmid in the pool - ex. the gRNA in a CRISPR library. In general, our QC scientists...

...is mediated by recruitment of MS2-functionalized gRNAs.  Find the yeast CRISPRa plasmids here Reduce ...

...interest is difficult to target specifically with a gRNA). Be sure to keep these techniques in mind when ...

...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 ...

...than relying on a two-component system dependent on gRNA design (SAM) or peptide design (SunTag). This streamlines...(SAM) Description SAM uses specially engineered sgRNAs to increase transcription. This is done through...

...transfect/inject cells with in vitro preassembled Cas9/gRNA complexes or with guide RNA- and Cas9-expression...

...edit to be made when multiple As are present in the gRNA spacer, as ADAR will preferentially edit an adenine...

...Addgene, including cDNA, barcoding, screening, and gRNA libraries. Most of the pooled libraries at Addgene...

...dedicated to expressing Cas9, while the other houses the gRNA and scaffold. If you don’t feel like doubling the...