CRISPR-SKIP: programmable gene splicing with single base editors

Genome Biol. 2018 Aug 15;19(1):107. doi: 10.1186/s13059-018-1482-5.

Abstract

CRISPR gene editing has revolutionized biomedicine and biotechnology by providing a simple means to engineer genes through targeted double-strand breaks in the genomic DNA of living cells. However, given the stochasticity of cellular DNA repair mechanisms and the potential for off-target mutations, technologies capable of introducing targeted changes with increased precision, such as single-base editors, are preferred. We present a versatile method termed CRISPR-SKIP that utilizes cytidine deaminase single-base editors to program 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.

Keywords: Alternative splicing; BRCA2; Base editing; CRISPR-Cas9; Exon skipping; Gene editing; Gene isoform; PIK3CA; RELA; Synthetic biology.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Base Pairing / genetics*
  • Base Sequence
  • Cell Line
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
  • Consensus Sequence / genetics
  • Exons / genetics
  • Gene Editing*
  • Genome
  • High-Throughput Nucleotide Sequencing
  • Humans
  • RNA Splice Sites / genetics

Substances

  • RNA Splice Sites