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Mol Cell Biol
2001 Jan 01;211:289-97. doi: 10.1128/MCB.21.1.289-297.2001.
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Stimulation of homologous recombination through targeted cleavage by chimeric nucleases.
Bibikova M
,
Carroll D
,
Segal DJ
,
Trautman JK
,
Smith J
,
Kim YG
,
Chandrasegaran S
.
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Chimeric nucleases that are hybrids between a nonspecific DNA cleavage domain and a zinc finger DNA recognition domain were tested for their ability to find and cleave their target sites in living cells. Both engineered DNA substrates and the nucleases were injected into Xenopus laevis oocyte nuclei, in which DNA cleavage and subsequent homologous recombination were observed. Specific cleavage required two inverted copies of the zinc finger recognition site in close proximity, reflecting the need for dimerization of the cleavage domain. Cleaved DNA molecules were activated for homologous recombination; in optimum conditions, essentially 100% of the substrate recombined, even though the DNA was assembled into chromatin. The original nuclease has an 18-amino-acid linker between the zinc finger and cleavage domains, and this enzyme cleaved in oocytes at paired sites separated by spacers in the range of 6 to 18 bp, with a rather sharp optimum at 8 bp. By shortening the linker, we found that the range of effective site separations could be narrowed significantly. With no intentional linker between the binding and cleavage domains, only binding sites exactly 6 bp apart supported efficient cleavage in oocytes. We also showed that two chimeric enzymes with different binding specificities could collaborate to stimulate recombination when their individual sites were appropriately placed. Because the recognition specificity of zinc fingers can be altered experimentally, this approach holds great promise for inducing targeted recombination in a variety of organisms.
Beerli,
Toward controlling gene expression at will: specific regulation of the erbB-2/HER-2 promoter by using polydactyl zinc finger proteins constructed from modular building blocks.
1998, Pubmed
Beerli,
Toward controlling gene expression at will: specific regulation of the erbB-2/HER-2 promoter by using polydactyl zinc finger proteins constructed from modular building blocks.
1998,
Pubmed
Bitinaite,
FokI dimerization is required for DNA cleavage.
1998,
Pubmed
Capecchi,
Altering the genome by homologous recombination.
1989,
Pubmed
Carroll,
Homologous genetic recombination in Xenopus: mechanism and implications for gene manipulation.
1996,
Pubmed
,
Xenbase
Carroll,
Efficient homologous recombination of linear DNA substrates after injection into Xenopus laevis oocytes.
1986,
Pubmed
,
Xenbase
Chandrasegaran,
Chimeric restriction enzymes: what is next?
1999,
Pubmed
Choo,
Toward a code for the interactions of zinc fingers with DNA: selection of randomized fingers displayed on phage.
1994,
Pubmed
Choo,
In vivo repression by a site-specific DNA-binding protein designed against an oncogenic sequence.
1994,
Pubmed
Choulika,
Induction of homologous recombination in mammalian chromosomes by using the I-SceI system of Saccharomyces cerevisiae.
1995,
Pubmed
Cohen-Tannoudji,
I-SceI-induced gene replacement at a natural locus in embryonic stem cells.
1998,
Pubmed
Desjarlais,
Toward rules relating zinc finger protein sequences and DNA binding site preferences.
1992,
Pubmed
Desjarlais,
Use of a zinc-finger consensus sequence framework and specificity rules to design specific DNA binding proteins.
1993,
Pubmed
Elliott,
Gene conversion tracts from double-strand break repair in mammalian cells.
1998,
Pubmed
Gargiulo,
Analysis of the chromatin assembled in germinal vesicles of Xenopus oocytes.
1983,
Pubmed
,
Xenbase
Greisman,
A general strategy for selecting high-affinity zinc finger proteins for diverse DNA target sites.
1997,
Pubmed
Huang,
Splase: a new class IIS zinc-finger restriction endonuclease with specificity for Sp1 binding sites.
1996,
Pubmed
Isalan,
Comprehensive DNA recognition through concerted interactions from adjacent zinc fingers.
1998,
Pubmed
Jamieson,
In vitro selection of zinc fingers with altered DNA-binding specificity.
1994,
Pubmed
Jasin,
Genetic manipulation of genomes with rare-cutting endonucleases.
1996,
Pubmed
Jeggo,
DNA breakage and repair.
1998,
Pubmed
Jeong-Yu,
Effect of terminal nonhomologies on homologous recombination in Xenopus laevis oocytes.
1992,
Pubmed
,
Xenbase
Kim,
Chimeric restriction endonuclease.
1994,
Pubmed
Kim,
Hybrid restriction enzymes: zinc finger fusions to Fok I cleavage domain.
1996,
Pubmed
Kim,
A 2.2 A resolution crystal structure of a designed zinc finger protein bound to DNA.
1996,
Pubmed
Kim,
Design of TATA box-binding protein/zinc finger fusions for targeted regulation of gene expression.
1997,
Pubmed
Kim,
Construction of a Z-DNA-specific restriction endonuclease.
1997,
Pubmed
Kim,
Site-specific cleavage of DNA-RNA hybrids by zinc finger/FokI cleavage domain fusions.
1997,
Pubmed
Kim,
Getting a handhold on DNA: design of poly-zinc finger proteins with femtomolar dissociation constants.
1998,
Pubmed
Kim,
Chimeric restriction enzyme: Gal4 fusion to FokI cleavage domain.
1998,
Pubmed
Koller,
Altering genes in animals by gene targeting.
1992,
Pubmed
Lieber,
The biochemistry and biological significance of nonhomologous DNA end joining: an essential repair process in multicellular eukaryotes.
1999,
Pubmed
Lin,
Model for homologous recombination during transfer of DNA into mouse L cells: role for DNA ends in the recombination process.
1984,
Pubmed
Liu,
Design of polydactyl zinc-finger proteins for unique addressing within complex genomes.
1997,
Pubmed
Maryon,
Degradation of linear DNA by a strand-specific exonuclease activity in Xenopus laevis oocytes.
1989,
Pubmed
,
Xenbase
Maryon,
Characterization of recombination intermediates from DNA injected into Xenopus laevis oocytes: evidence for a nonconservative mechanism of homologous recombination.
1991,
Pubmed
,
Xenbase
Nahon,
Targeting a truncated Ho-endonuclease of yeast to novel DNA sites with foreign zinc fingers.
1998,
Pubmed
Osman,
Double-strand break-induced recombination in eukaryotes.
1998,
Pubmed
Ozenberger,
A unique pathway of double-strand break repair operates in tandemly repeated genes.
1991,
Pubmed
Plessis,
Site-specific recombination determined by I-SceI, a mitochondrial group I intron-encoded endonuclease expressed in the yeast nucleus.
1992,
Pubmed
Puchta,
Homologous recombination in plant cells is enhanced by in vivo induction of double strand breaks into DNA by a site-specific endonuclease.
1993,
Pubmed
Pâques,
Multiple pathways of recombination induced by double-strand breaks in Saccharomyces cerevisiae.
1999,
Pubmed
Rebar,
Zinc finger phage: affinity selection of fingers with new DNA-binding specificities.
1994,
Pubmed
Richardson,
Double-strand break repair by interchromosomal recombination: suppression of chromosomal translocations.
1998,
Pubmed
Rong,
Gene targeting by homologous recombination in Drosophila.
2000,
Pubmed
Rouet,
Introduction of double-strand breaks into the genome of mouse cells by expression of a rare-cutting endonuclease.
1994,
Pubmed
Rudin,
Efficient repair of HO-induced chromosomal breaks in Saccharomyces cerevisiae by recombination between flanking homologous sequences.
1988,
Pubmed
Segal,
Endonuclease-induced, targeted homologous extrachromosomal recombination in Xenopus oocytes.
1995,
Pubmed
,
Xenbase
Segal,
Toward controlling gene expression at will: selection and design of zinc finger domains recognizing each of the 5'-GNN-3' DNA target sequences.
1999,
Pubmed
Shi,
Specific DNA-RNA hybrid binding by zinc finger proteins.
1995,
Pubmed
Smith,
A detailed study of the substrate specificity of a chimeric restriction enzyme.
1999,
Pubmed
Smith,
Requirements for double-strand cleavage by chimeric restriction enzymes with zinc finger DNA-recognition domains.
2000,
Pubmed
Thomas,
High frequency targeting of genes to specific sites in the mammalian genome.
1986,
Pubmed
Wah,
Structure of the multimodular endonuclease FokI bound to DNA.
1997,
Pubmed
Wah,
Structure of FokI has implications for DNA cleavage.
1998,
Pubmed
Widom,
Structure, dynamics, and function of chromatin in vitro.
1998,
Pubmed
Wolfe,
Analysis of zinc fingers optimized via phage display: evaluating the utility of a recognition code.
1999,
Pubmed
Wu,
Building zinc fingers by selection: toward a therapeutic application.
1995,
Pubmed
Zheng,
Gene targeting in normal and amplified cell lines.
1990,
Pubmed
