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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,
Efficient homologous recombination of linear DNA substrates after injection into Xenopus laevis oocytes.
1986,
Pubmed
,
Xenbase Carroll,
Homologous genetic recombination in Xenopus: mechanism and implications for gene manipulation.
1996,
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,
Use of a zinc-finger consensus sequence framework and specificity rules to design specific DNA binding proteins.
1993,
Pubmed Desjarlais,
Toward rules relating zinc finger protein sequences and DNA binding site preferences.
1992,
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,
Construction of a Z-DNA-specific restriction endonuclease.
1997,
Pubmed Kim,
Chimeric restriction endonuclease.
1994,
Pubmed Kim,
Hybrid restriction enzymes: zinc finger fusions to Fok I cleavage domain.
1996,
Pubmed Kim,
Chimeric restriction enzyme: Gal4 fusion to FokI cleavage domain.
1998,
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,
Getting a handhold on DNA: design of poly-zinc finger proteins with femtomolar dissociation constants.
1998,
Pubmed Kim,
Site-specific cleavage of DNA-RNA hybrids by zinc finger/FokI cleavage domain fusions.
1997,
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,
Characterization of recombination intermediates from DNA injected into Xenopus laevis oocytes: evidence for a nonconservative mechanism of homologous recombination.
1991,
Pubmed
,
Xenbase Maryon,
Degradation of linear DNA by a strand-specific exonuclease activity in Xenopus laevis oocytes.
1989,
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 Pâques,
Multiple pathways of recombination induced by double-strand breaks in Saccharomyces cerevisiae.
1999,
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 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
