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Nuclease‐mediated genome editing: At the front‐line of functional genomics technology
Author(s) -
Sakuma Tetsushi,
Woltjen Knut
Publication year - 2014
Publication title -
development, growth and differentiation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.864
H-Index - 66
eISSN - 1440-169X
pISSN - 0012-1592
DOI - 10.1111/dgd.12111
Subject(s) - transcription activator like effector nuclease , genome editing , zinc finger nuclease , crispr , biology , genome engineering , nuclease , homology directed repair , computational biology , genetics , cas9 , genome , functional genomics , dna , genomics , dna repair , gene , nucleotide excision repair
Genome editing with engineered endonucleases is rapidly becoming a staple method in developmental biology studies. Engineered nucleases permit random or designed genomic modification at precise loci through the stimulation of endogenous double‐strand break repair. Homology‐directed repair following targeted DNA damage is mediated by co‐introduction of a custom repair template, allowing the derivation of knock‐out and knock‐in alleles in animal models previously refractory to classic gene targeting procedures. Currently there are three main types of customizable site‐specific nucleases delineated by the source mechanism of DNA binding that guides nuclease activity to a genomic target: zinc‐finger nucleases ( ZFN s), transcription activator‐like effector nucleases ( TALEN s), and clustered regularly interspaced short palindromic repeats ( CRISPR ). Among these genome engineering tools, characteristics such as the ease of design and construction, mechanism of inducing DNA damage, and DNA sequence specificity all differ, making their application complementary. By understanding the advantages and disadvantages of each method, one may make the best choice for their particular purpose.