Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease.
basic_science · Level V
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- Record sourced from PubMed, PMID 26436575.
- Also identified by PMC identifier 4747789.
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Abstract
We have developed a CRISPR-based method that uses catalytically active Cas9 and distinct single guide (sgRNA) constructs to knock out and activate different genes in the same cell. These sgRNAs, with 14- to 15-bp target sequences and MS2 binding loops, can activate gene expression using an active Streptococcus pyogenes Cas9 nuclease, without inducing double-stranded breaks. We use these 'dead RNAs' to perform orthogonal gene knockout and transcriptional activation in human cells.
Medical subject headings
- Bacterial Proteins
- Endonucleases
- Gene Knockout Techniques
- RNA, Guide, CRISPR-Cas Systems