title: Precision genome engineering with programmable DNA-nicking enzymes.
abstract: Zinc finger nucleases (ZFNs) are powerful tools of genome engineering but are limited by their inevitable reliance on error-prone nonhomologous end-joining (NHEJ) repair of DNA double-strand breaks (DSBs), which gives rise to randomly generated, unwanted small insertions or deletions (indels) at both on-target and off-target sites. Here, we present programmable DNA-nicking enzymes (nickases) that produce single-strand breaks (SSBs) or nicks, instead of DSBs, which are repaired by error-free homologous recombination (HR) rather than mutagenic NHEJ. Unlike their corresponding nucleases, zinc finger nickases allow site-specific genome modifications only at the on-target site, without the induction of unwanted indels. We propose that programmable nickases will be of broad utility in research, medicine, and biotechnology, enabling precision genome engineering in any cell or organism.
authors.affiliation: National Creative Research Initiatives Center for Genome Engineering and Department of Chemistry, Seoul National University, 599 Gwanak-ro, Gwanak-gu, Seoul, South Korea.
authors.forename: Eunji
authors.initials: E
authors.lastname: Kim
authors.forename: Sojung
authors.initials: S
authors.lastname: Kim
authors.forename: Duk Hyoung
authors.initials: DH
authors.lastname: Kim
authors.forename: Beom-Soon
authors.initials: BS
authors.lastname: Choi
authors.forename: Ik-Young
authors.initials: IY
authors.lastname: Choi
authors.forename: Jin-Soo
authors.initials: JS
authors.lastname: Kim
mesh_terms: D003001:Cloning, Molecular; D053903:DNA Breaks, Double-Stranded; D053904:DNA Breaks, Single-Stranded; D059766:DNA End-Joining Repair; D003850:Deoxyribonuclease I / Q000235:genetics / Q000378:metabolism*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000235:genetics / Q000378:metabolism*; D018390:Gene Targeting / Q000379:methods; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D015894:Genome, Human; D057809:HEK293 Cells; D006801:Humans; D054643:INDEL Mutation; D012680:Sensitivity and Specificity; D016335:Zinc Fingers
chemical_list: C035042:endodeoxyribonuclease FokI; D003850:Deoxyribonuclease I; D015252:Deoxyribonucleases, Type II Site-Specific