CRISPR-SKIP: Programmable Gene Splicing with Single Base Editors [gDNA]. CRISPR-SKIP: Programmable Gene Splicing with Single Base Editors [gDNA]

CRISPR gene editing has revolutionized biomedicine and biotechnology by providing a simple means to engineer genes in vivo by introducing mutations at target sites in the genomic DNA of living cells. However, given the stochasticity of cellular DNA repair mechanisms and the potential for introducing mutations at off-target sites, technologies capable of introducing targeted changes with increased precision, such as cytidine deaminase single-base editors, are preferred. We here present a versatile method termed CRISPR-SKIP that utilizes cytidine deaminase single-base editors to program de-novo exon skipping by mutating target DNA bases within splice acceptor sites. Given its simplicity and precision, CRISPR-SKIP will be broadly applicable in gene therapy and synthetic biology. Overall design: Four exons were targeted by CRISPR-SKIP. For each exon, genomic DNA at the splice acceptor of two control samples and two treated samples were sequenced. Rate of G to A conversion of intronic flanking G was quantified.