Zinc finger nucleases: custom-designed molecular scissors for genome engineering of plant and mammalian cells.
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.
Durai
Sundar
S
Department of Environmental Health Sciences, The Johns Hopkins University Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, MD 21205-2179, USA.
Mani
Mala
M
Kandavelou
Karthikeyan
K
Wu
Joy
J
Porteus
Matthew H
MH
Chandrasegaran
Srinivasan
S
D000818:Animals; D020134:Catalytic Domain; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000737:chemistry*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry*; D018390:Gene Targeting; D005818:Genetic Engineering; D015894:Genome, Human; D018745:Genome, Plant; D023281:Genomics; D006801:Humans; D016335:Zinc Fingers
D004268:DNA-Binding Proteins; C035042:endodeoxyribonuclease FokI; D015252:Deoxyribonucleases, Type II Site-Specific