The democratization of gene editing: Insights from site-specific cleavage and double-strand break repair.
DNA double-strand breaks (DSBs) are dangerous lesions that if not properly repaired can lead to genomic change or cell death. Organisms have developed several pathways and have many factors devoted to repairing DSBs, which broadly occurs by homologous recombination, which relies on an identical or homologous sequence to template repair, or nonhomologous end-joining. Much of our understanding of these repair mechanisms has come from the study of induced DNA cleavage by site-specific endonucleases. In addition to their biological role, these cellular pathways can be co-opted for gene editing to study gene function or for gene therapy or other applications. While the first gene editing experiments were done more than 20 years ago, the recent discovery of RNA-guided endonucleases has simplified approaches developed over the years to make gene editing an approach that is available to the entire biomedical research community. Here, we review DSB repair mechanisms and site-specific cleavage systems that have provided insight into these mechanisms and led to the current gene editing revolution.
Developmental Biology Program, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10065, USA. Electronic address: m-jasin@ski.mskcc.org.
Maria
M
Jasin
Department of Biology and Rosenstiel Basic Medical Sciences Research Center, Brandeis University, 02454-9110, USA. Electronic address: haber@brandeis.edu.
James E
JE
Haber
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D000076987:CRISPR-Associated Protein 9; D064113:CRISPR-Cas Systems; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D053903:DNA Breaks, Double-Stranded; D059766:DNA End-Joining Repair; D053843:DNA Mismatch Repair; D015252:Deoxyribonucleases, Type II Site-Specific / Q000235:genetics / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000235:genetics / Q000378:metabolism; D004720:Endonucleases / Q000235:genetics / Q000378:metabolism; D000072669:Gene Editing / Q000379:methods*; D006801:Humans; D012194:RNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism; D059767:Recombinational DNA Repair; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D000069896:Transcription Activator-Like Effector Nucleases / Q000235:genetics / Q000378:metabolism; D016335:Zinc Fingers / Q000235:genetics
D001426:Bacterial Proteins; D029701:Saccharomyces cerevisiae Proteins; D004247:DNA; D012194:RNA-Directed DNA Polymerase; C454879:maturase I-AniI; D000076987:CRISPR-Associated Protein 9; C000600049:Cas9 protein, Francisella novicida; D004706:Endodeoxyribonucleases; D004720:Endonucleases; C064579:I-Ppo endonuclease; D000069896:Transcription Activator-Like Effector Nucleases; C037725:SCEI protein, S cerevisiae; D015252:Deoxyribonucleases, Type II Site-Specific
CRISPR-Cas9; Double-strand break repair; Gene editing; HO endonuclease; Homologous recombination; Homology-directed repair; I-SceI endonuclease; Nonhomologous end-joining