摘要
Zinc-finger nucleases and transcription activator-like effector nucleases are novel gene-editing platformscontributing to redefine the boundaries of modern biological research. They are composed of a non-specificcleavage domain and a tailor made DNA-binding module, which enables a broad range of genetic modifications byinducing efficient DNA double-strand breaks at desired loci. Among other remarkable uses, these nucleases havebeen employed to produce gene knockouts in mid-size and large animals, such as rabbits and pigs, respectively.This approach is cost effective, relatively quick, and can produce invaluable models for human disease studies,biotechnology or agricultural purposes. Here we describe a protocol for the efficient generation of knockout rabbitsusing transcription activator-like effector nucleases, and a perspective of the field.
基金
Work on this topic in the authors’laboratories is supported by grants from:the Strategic Priority Research Program of the Chinese Academy of Sciences(number XDA01020106)
the Ministry of Science and Technology of China 973 program(2011CB965200)
the National Natural Science Foundation of China(81261130317)to MAE
the Bureau of Science,Technology and Information of Guangzhou Municipality(2012 J5100040)to MAE and JF
grants 2010U1-E00811-5 and ZNGI-2011-010 from the Guangzhou Municipality and the Chinese Academy of Sciences,respectively,to LL.