title: Site-specific cleavage of DNA-RNA hybrids by zinc finger/FokI cleavage domain fusions.
abstract: Zinc-finger proteins of the Cys2His2 type bind DNA-RNA hybrids with affinities comparable to those for DNA duplexes. Such zinc-finger proteins were converted into site-specific cleaving enzymes by fusing them to the FokI cleavage domain. The fusion proteins are active and under optimal conditions cleave DNA duplexes in a sequence-specific manner. These fusions also exhibit site-specific cleavage of the DNA strand within DNA-RNA hybrids albeit at a lower efficiency (approximately 50-fold) compared to the cleavage of the DNA duplexes. These engineered endonucleases represent the first of their kind in terms of their DNA-RNA cleavage properties, and they may have important biological applications.
authors.affiliation: Department of Environmental Health Sciences, The Johns Hopkins University, School of Hygiene and Public Health, Baltimore, MD 21205-2179, USA.
authors.forename: Y G
authors.initials: YG
authors.lastname: Kim
authors.forename: Y
authors.initials: Y
authors.lastname: Shi
authors.forename: J M
authors.initials: JM
authors.lastname: Berg
authors.forename: S
authors.initials: S
authors.lastname: Chandrasegaran
mesh_terms: D001483:Base Sequence; D001665:Binding Sites; D004247:DNA / Q000378:metabolism*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D012313:RNA / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D016335:Zinc Fingers
chemical_list: D011993:Recombinant Fusion Proteins; D012313:RNA; D004247:DNA; C035042:endodeoxyribonuclease FokI; D015252:Deoxyribonucleases, Type II Site-Specific