CRISPR-SKIP: programmable gene splicing with single base editors

• Published in: Genome Biology Vol. 19; no. 1; p. 107
• Main Authors: Gapinske, Michael, Luu, Alan, Winter, Jackson, Woods, Wendy S., Kostan, Kurt A., Shiva, Nikhil, Song, Jun S., Perez-Pinera, Pablo
• Format: Journal Article
• Language: English
• Published: London BioMed Central 15.08.2018; Springer Nature B.V; BMC
• Subjects: Alternative splicing; Animal Genetics and Genomics; Base editing; Base Pairing - genetics; Base Sequence; Bioinformatics; Biomedical and Life Sciences; Biotechnology; Cell Line; Cloning; Clustered Regularly Interspaced Short Palindromic Repeats - genetics; Consensus Sequence - genetics; CRISPR; CRISPR-Cas9; Cytidine deaminase; Deoxyribonucleic acid; DNA; DNA repair; Efficiency; Evolutionary Biology; Exon skipping; exons; Exons - genetics; Gene Editing; Gene expression; Gene therapy; Genome; Genomes; High-Throughput Nucleotide Sequencing; Human Genetics; Humans; Insights from Genome Editing; Life Sciences; medicine; Method; Microbial Genetics and Genomics; Muscular dystrophy; Mutation; Penicillin; Plant Genetics and Genomics; Plasmids; Proteins; RNA Splice Sites - genetics; Splicing; Stochasticity; Synthetic biology; Alternative splicing; Gene isoform; Gene editing; Base editing; Exon skipping; CRISPR-Cas9; Synthetic biology; PIK3CA; BRCA2; RELA
• ISSN: 1474-760X; 1474-7596; 1474-760X
• DOI: 10.1186/s13059-018-1482-5

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.