Mutations were targeted to the Hprt locus of mouse embryo-derived stem cells by using 22 different sequence replacement and sequence insertion vectors. The targeting frequency was examined at two sites within the Hprt locus as a function of the extent of homology between the targeting vector and the target locus. The targeting frequency was also compared by using vectors prepared from isogenic and nonisogenic DNA sources. With one exception, all of the vectors showed the same exponential dependence of targeting efficiency on the extent of homology between the targeting vector and the target locus. This was true regardless of whether they were sequence replacement or sequence insertion vectors, whether they were directed toward either of the two different sites within the Hprt locus, or whether they were prepared from isogenic or nonisogenic DNA sources. Vectors prepared from isogenic DNA targeted four to five times more efficiently than did the corresponding vectors prepared from nonisogenic DNA. The single case of unexpectedly low targeting efficiency involved one of the vectors prepared from nonisogenic DNA and could be attributed to an unfavorable distribution of heterology between the Hprt sequences present in the targeting...Continue Reading
The barrier to recombination between Escherichia coli and Salmonella typhimurium is disrupted in mismatch-repair mutants
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Trypanosoma brucei homologous recombination is dependent on substrate length and homology, though displays a differential dependence on mismatch repair as substrate length decreases
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Simple and straightforward construction of a mouse gene targeting vector using in vitro transposition reactions
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A Functional chromatin domain does not resist X chromosome inactivation: silencing of cLys correlates with methylation of a dual promoter-replication origin
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Campylobacter fetus uses multiple loci for DNA inversion within the 5' conserved regions of sap homologs
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Targeting vector configuration and method of gene transfer influence targeted correction of the APRT gene in Chinese hamster ovary cells
Long targeting arms do not increase the efficiency of homologous recombination in the beta-globin locus of murine embryonic stem cells.
Stimulation of oligonucleotide-directed gene correction by Redβ expression and MSH2 depletion in human HT1080 cells
Optimization of the production of knock-in alleles by CRISPR/Cas9 microinjection into the mouse zygote
Expanding the editable genome and CRISPR-Cas9 versatility using DNA cutting-free gene targeting based on in trans paired nicking
Optimizing CRISPR/Cas9 technology for precise correction of the Fgfr3-G374R mutation in achondroplasia in mice.
Investigation of the molecular biology underlying the pronounced high gene targeting frequency at the Myh9 gene locus in mouse embryonic stem cells
Mouse model of X-linked chronic granulomatous disease, an inherited defect in phagocyte superoxide production
Cloning of the murine TROP2 gene: conservation of a PIP2-binding sequence in the cytoplasmic domain of TROP-2
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Site-specific gene targeting in mouse embryonic stem cells with intact bacterial artificial chromosomes
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A protocol for constructing gene targeting vectors: generating knockout mice for the cadherin family and beyond
Homologous recombination in human embryonic stem cells: a tool for advancing cell therapy and understanding and treating human disease
Multiple copies of a linear donor fragment released in situ from a vector improve the efficiency of zinc-finger nuclease-mediated genome editing
1,5-isoquinolinediol increases the frequency of gene targeting by homologous recombination in mouse fibroblasts
Correction of mutant Fanconi anemia gene by homologous recombination in human hematopoietic cells using adeno-associated virus vector
Simple one-week method to construct gene-targeting vectors: application to production of human knockout cell lines
Introduction of hereditary disease-associated mutations into the beta-amyloid precursor protein gene of mouse embryonic stem cells: a comparison of homologous recombination methods
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