TY - JOUR
T1 - Zinc finger nucleases
T2 - Custom-designed molecular scissors for genome engineering of plant and mammalian cells
AU - Durai, Sundar
AU - Mani, Mala
AU - Kandavelou, Karthikeyan
AU - Wu, Joy
AU - Porteus, Matthew H.
AU - Chandrasegaran, Srinivasan
N1 - Funding Information:
This work was funded by a grant from National Institutes of Health (GM 53923). We also thank Environmental Health Sciences Center Core Facility (supported by grant ES 03819) for synthesis of oligonucleotides. K.K. was supported for a year by the Johns Hopkins University Center for AIDS Research (CFAR; Grant # P30 AI42855) grant. Funding to pay the Open Access publication charges for this article was provided by JHU.
PY - 2005
Y1 - 2005
N2 - 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.
AB - 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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U2 - 10.1093/nar/gki912
DO - 10.1093/nar/gki912
M3 - Article
C2 - 16251401
AN - SCOPUS:27744530147
SN - 0305-1048
VL - 33
SP - 5978
EP - 5990
JO - Nucleic acids research
JF - Nucleic acids research
IS - 18
ER -