Zinc finger nucleases: custom-designed molecular scissors for genome engineering of plant and mammalian cells

Zinc finger nucleases: custom-designed molecular scissors for genome engineering of plant and mammalian cells

Sundar Durai, Mala Mani, Karthikeyan Kandavelou, Joy Wu1, Matthew H. Porteus and Srinivasan Chandrasegaran

Nucleic Acids Research Volume33, Issue18Pp. 5978-5990 doi:10.1093/nar/gki912


Custom-designed zinc finger nucleases (ZFNs), proteins designed to cut at specific DNA sequences, are becoming powerful tools in gene targeting—the process of replacing a gene within a genome by homologous recombination (HR). ZFNs that combine the non-specific cleavage domain (N) of FokI endonuclease with zinc finger proteins (ZFPs) offer a general way to deliver a site-specific double-strand break (DSB) to the genome. The development of ZFN-mediated gene targeting provides molecular biologists with the ability to site-specifically and permanently modify plant and mammalian genomes including the human genome via homology-directed repair of a targeted genomic DSB. The creation of designer ZFNs that cleave DNA at a pre-determined site depends on the reliable creation of ZFPs that can specifically recognize the chosen target site within a genome. The (Cys2His2) ZFPs offer the best framework for developing custom ZFN molecules with new sequence-specificities. Here, we explore the different approaches for generating the desired custom ZFNs with high sequence-specificity and affinity. We also discuss the potential of ZFN-mediated gene targeting for ‘directed mutagenesis’ and targeted ‘gene editing’ of the plant and mammalian genome as well as the potential of ZFN-based strategies as a form of gene therapy for human therapeutics in the future.

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  1. #1 par peptide 3332 le janvier 19, 2011 - 9:16

    very nice article .. thanks for sharing

  1. Tweets that mention Zinc finger nucleases: custom-designed molecular scissors for genome engineering of plant and mammalian cells | Coffee and Sci(ence) -- Topsy.com

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