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

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

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish (US)
Pages (from-to)5978-5990
Number of pages13
JournalNucleic acids research
Volume33
Issue number18
DOIs
StatePublished - 2005

ASJC Scopus subject areas

  • Genetics

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