期刊
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
卷 335, 期 2, 页码 447-457出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bbrc.2005.07.089
关键词
chimeric nucleases; gene targeting; homologous recombination; non-homologous end joining; protein design; protein engineering; sequence-specific cleavage
资金
- NIAID NIH HHS [P30 AI42855] Funding Source: Medline
- NIGMS NIH HHS [GM 53923] Funding Source: Medline
Zinc finger nuclease (ZFN)-mediated gene targeting is rapidly becoming a powerful too] for gene editing and directed mutagenesis of plant and mammalian genomes including the human genome. ZFN-mediated gene targeting provides molecular biologists with the ability to site-specifically manipulate and permanently modify plant and mammalian genomes. Facile production of ZFNs and rapid characterization of their in vitro sequence-specific cleavage properties are a pre-requisite before ZFN-mediated gene targeting can become an efficient and effective practical tool for widespread use in biotechnology. Here, we report the design, engineering, and rapid in vitro characterization of ZFNs that target specific endogenous sequences within two mouse genes (mTYR and mCFTR), and two human genes (hCCR5 and hDMPK), respectively. These engineered ZFNs recognize their respective cognate DNA sites encoded in a plasmid substrate in a sequence-specific manner and, as expected, they induce a double-strand break at the chosen target site. (c) 2005 Elsevier Inc. All rights reserved.
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