Designing Guide-RNA for Generating Premature Stop Codons for Gene Knockout Using CRISPR-BETS.
Methods in molecular biology (Clifton, N.J.), 2653: 95-105 PMID 36995621 DOI 10.1007/978-1-0716-3131-7_6
Abstract
Cytosine base editors (CBEs) accurately modify target sites by mediating a C to T change (or a G to A change on the opposite strand). This allows us to install premature stop codons for gene knockout. However, highly specific sgRNAs (single-guide RNAs) are necessary for the CRISPR-Cas nuclease to work efficiently. In this study, we introduce a method of designing highly specific gRNA to generate premature stop codons and knock out a gene using CRISPR-BETS software.