Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
???displayArticle.abstract???
Homologous recombination in gene targeting in most organisms occurs by an inefficient mechanism. Inducing a double-strand break in the chromosomal target may increase this efficiency by allowing recombination to proceed by the highly efficient single-strand annealing mechanism. A gene targeting experiment was modeled in Xenopus oocytes by using a circular plasmid to mimic the chromosomal target site and a homologous linear molecule (pick-up fragment or PUF) as an analogue of the vector DNA. When those two molecules were simply injected together, no recombination was observed. In contrast, when the circular plasmid was cleaved in vivo by injection of the site-specific endonuclease, I-Sce I, relatively efficient intermolecular recombination occurred, involving up to 17% of the cleaved molecules. Recombination was dependent on the stability of the PUF; product yield was increased by using longer fragments and by injecting larger amounts of linear DNA, both of which increased the lifetime of the PUF in the oocytes. These results demonstrate that in vivo double-strand breaks can induce homologous recombination of reluctant substrates and may be useful in augmenting the efficiency of gene targeting.
Capecchi,
Altering the genome by homologous recombination.
1989, Pubmed
Capecchi,
Altering the genome by homologous recombination.
1989,
Pubmed Capecchi,
Gene targeting. How efficient can you get?
1990,
Pubmed Carroll,
Efficient homologous recombination of linear DNA substrates after injection into Xenopus laevis oocytes.
1986,
Pubmed
,
Xenbase Carroll,
Isolated clusters of paired tandemly repeated sequences in the Xenopus laevis genome.
1984,
Pubmed
,
Xenbase Church,
Genomic sequencing.
1984,
Pubmed Colleaux,
Recognition and cleavage site of the intron-encoded omega transposase.
1988,
Pubmed Deng,
Reexamination of gene targeting frequency as a function of the extent of homology between the targeting vector and the target locus.
1992,
Pubmed Feinberg,
A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.
1983,
Pubmed Gargiulo,
Analysis of the chromatin assembled in germinal vesicles of Xenopus oocytes.
1983,
Pubmed
,
Xenbase Garrett,
Tx1: a transposable element from Xenopus laevis with some unusual properties.
1986,
Pubmed
,
Xenbase Gloor,
Targeted gene replacement in Drosophila via P element-induced gap repair.
1991,
Pubmed Hasty,
The role and fate of DNA ends for homologous recombination in embryonic stem cells.
1992,
Pubmed Jasin,
Homologous integration in mammalian cells without target gene selection.
1988,
Pubmed Jeong-Yu,
Effect of terminal nonhomologies on homologous recombination in Xenopus laevis oocytes.
1992,
Pubmed
,
Xenbase Jeong-Yu,
Test of the double-strand-break repair model of recombination in Xenopus laevis oocytes.
1992,
Pubmed
,
Xenbase Koller,
Altering genes in animals by gene targeting.
1992,
Pubmed Lehman,
Homologous and illegitimate recombination in developing Xenopus oocytes and eggs.
1993,
Pubmed
,
Xenbase Lin,
Model for homologous recombination during transfer of DNA into mouse L cells: role for DNA ends in the recombination process.
1984,
Pubmed Mansour,
Disruption of the proto-oncogene int-2 in mouse embryo-derived stem cells: a general strategy for targeting mutations to non-selectable genes.
1988,
Pubmed Maryon,
Degradation of linear DNA by a strand-specific exonuclease activity in Xenopus laevis oocytes.
1989,
Pubmed
,
Xenbase Maryon,
Involvement of single-stranded tails in homologous recombination of DNA injected into Xenopus laevis oocyte nuclei.
1991,
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 Miller,
Template structural requirements for transcription in vivo by RNA polymerase II.
1982,
Pubmed
,
Xenbase Ozenberger,
A unique pathway of double-strand break repair operates in tandemly repeated genes.
1991,
Pubmed Pei,
Site-specific cleavage of duplex DNA by a semisynthetic nuclease via triple-helix formation.
1990,
Pubmed Pennington,
Gene targeting in Chinese hamster ovary cells is conservative.
1991,
Pubmed Perrin,
Asymmetrical recognition and activity of the I-SceI endonuclease on its site and on intron-exon junctions.
1993,
Pubmed Perrouault,
Sequence-specific artificial photo-induced endonucleases based on triple helix-forming oligonucleotides.
1990,
Pubmed Plasterk,
Targeted alterations of the Caenorhabditis elegans genome by transgene instructed DNA double strand break repair following Tc1 excision.
1992,
Pubmed Plessis,
Site-specific recombination determined by I-SceI, a mitochondrial group I intron-encoded endonuclease expressed in the yeast nucleus.
1992,
Pubmed Pont-Kingdon,
Intermediates in extrachromosomal homologous recombination in Xenopus laevis oocytes: characterization by electron microscopy.
1993,
Pubmed
,
Xenbase 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 Rouet,
Expression of a site-specific endonuclease stimulates homologous recombination in mammalian cells.
1994,
Pubmed Rudin,
Genetic and physical analysis of double-strand break repair and recombination in Saccharomyces cerevisiae.
1989,
Pubmed Rudin,
Efficient repair of HO-induced chromosomal breaks in Saccharomyces cerevisiae by recombination between flanking homologous sequences.
1988,
Pubmed Strobel,
Site-specific cleavage of a yeast chromosome by oligonucleotide-directed triple-helix formation.
1990,
Pubmed Sweigert,
Repair and recombination of X-irradiated plasmids in Xenopus laevis oocytes.
1990,
Pubmed
,
Xenbase Thomas,
Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells.
1987,
Pubmed Thomas,
High frequency targeting of genes to specific sites in the mammalian genome.
1986,
Pubmed Wyllie,
Selective DNA conservation and chromatin assembly after injection of SV40 DNA into Xenopus oocytes.
1978,
Pubmed
,
Xenbase Zheng,
Gene targeting in normal and amplified cell lines.
1990,
Pubmed
