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J Cell Physiol. 2016 Nov;231(11):2380-92. doi: 10.1002/jcp.25367. Epub 2016 Mar 15.

Programmable Site-Specific Nucleases for Targeted Genome Engineering in Higher Eukaryotes.

Author information

1
Department of Biotechnology, Molecular Biology Laboratory, MS Swaminathan Research Foundation, Taramani, Chennai, India.

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.

PMID:
26945523
DOI:
10.1002/jcp.25367
[Indexed for MEDLINE]

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