What is a single guide RNA, and what role does it play in CRISPR-Cas genome editing...

What is a single guide RNA, and what role does it play in CRISPR-Cas genome editing in eukaryotic cells?

Expert Solution

A version of the naturally occurring two-piece guide RNA complex engineered into a single, continuous sequence. The simplified single-guide RNA is used to direct the Cas9 protein to bind and cleave a particular DNA sequence for genome editing.

The role of guide RNAs in CRISPR Cas-9 gene knockout

The S. pyogenes Cas9 nuclease can be programmed by a guide RNA to create double-strand breaks (DSB) at a particular genomic region. Imperfect repair of the DSB can lead to loss of function of the targeted gene. Although most guide RNA target sites result in some level of DNA editing (insertions or deletions), not all result in editing that causes a functional gene disruption or gene knockout. Therefore, it is necessary to consider various factors, including the location of the CRISPR-Cas9 target sequence within the gene, when designing a guide RNA for gene knockout.

gladiator answered 2 years ago

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