title: Programmable Site-Specific Nucleases for Targeted Genome Engineering in Higher Eukaryotes.
abstract: Recent advances in the targeted genome engineering enable molecular biologists to generate sequence specific modifications with greater efficiency and higher specificity in complex eukaryotic genomes. Programmable site-specific DNA cleavage reagents and cellular DNA repair mechanisms have made this possible. These reagents have become powerful tools for delivering a site-specific genomic double-strand break (DSB) at the desired chromosomal locus, which produces sequence alterations through error-prone non-homologous end joining (NHEJ) resulting in gene inactivations/knockouts. Alternatively, the DSB can be repaired through homology-directed repair (HDR) using a donor DNA template, which leads to the introduction of desired sequence modifications at the predetermined site. Here, we summarize the role of three classes of nucleases; zinc finger nucleases (ZFNs), transcription activator like effector nucleases (TALENs), and clustered regularly interspaced palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9) system in achieving targeted genome modifications. Further, we discuss the progress towards the applications of programmable site-specific nucleases (SSNs) in treating human diseases and other biological applications in economically important higher eukaryotic organisms such as plants and livestock. J. Cell. Physiol. 231: 2380-2392, 2016. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Department of Biotechnology, Molecular Biology Laboratory, MS Swaminathan Research Foundation, Taramani, Chennai, India.
authors.forename: Ganesan
authors.initials: G
authors.lastname: Govindan
authors.affiliation: Department of Biotechnology, Molecular Biology Laboratory, MS Swaminathan Research Foundation, Taramani, Chennai, India.
authors.forename: Sivaprakash
authors.initials: S
authors.lastname: Ramalingam
mesh_terms: D000818:Animals; D001483:Base Sequence; D004720:Endonucleases / Q000378:metabolism*; D056890:Eukaryota / Q000235:genetics*; D018390:Gene Targeting; D005818:Genetic Engineering / Q000379:methods*; D016678:Genome; D006801:Humans
chemical_list: D004720:Endonucleases