ArticleMolecular and cellular biology2001
The mechanism of mammalian gene replacement is consistent with the formation of long regions of heteroduplex DNA associated with two crossing-over events.
Article in Molecular and cellular biology, 2001. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed.
- Trial
- Molecular characterization of precise in vivo targeted gene integration in human cells using AAVHSC15.PloS one · 2020Article
- Gene Editing with Helper-Dependent Adenovirus Can Efficiently Introduce Multiple Changes Simultaneously over a Large Genomic Region.Molecular therapy. Nucleic acids · 2017Article
- Mechanisms of precise genome editing using oligonucleotide donors.Genome research · 2017Article
- Immunogenomic engineering of a plug-and-(dis)play hybridoma platform.Nature communications · 2016Article
- The mechanism of gene targeting in human somatic cells.PLoS genetics · 2014Article
- Effects of varying gene targeting parameters on processing of recombination intermediates by ERCC1-XPF.DNA repair · 2011Article
- Single-stranded heteroduplex intermediates in lambda Red homologous recombination.BMC molecular biology · 2010Article
- A caveat in mouse genetic engineering: ectopic gene targeting in ES cells by bidirectional extension of the homology arms of a gene replacement vector carrying human PARP-1.Transgenic research · 2009Article
- Efficient transfer of base changes from a vector to the rice genome by homologous recombination: involvement of heteroduplex formation and mismatch correction.Nucleic acids research · 2008Article
- Interchromosomal crossover in human cells is associated with long gene conversion tracts.Molecular and cellular biology · 2007Article
- Trypanosoma brucei homologous recombination is dependent on substrate length and homology, though displays a differential dependence on mismatch repair as substrate length decreases.Nucleic acids research · 2007Article
- Opposing roles for DNA structure-specific proteins Rad1, Msh2, Msh3, and Sgs1 in yeast gene targeting.The EMBO journal · 2005Article
- Gene targeting in yeast is initiated by two independent strand invasions.Proceedings of the National Academy of Sciences of the United States of America · 2004Article
- Gene repeat expansion and contraction by spontaneous intrachromosomal homologous recombination in mammalian cells.Nucleic acids research · 2004Article
- Repair of double-strand breaks by homologous recombination in mismatch repair-defective mammalian cells.Molecular and cellular biology · 2001Article
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3 authors.
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Abstract
In this study, the mechanism of mammalian gene replacement was investigated. The system is based on detecting homologous recombination between transferred vector DNA and the haploid, chromosomal immunoglobulin mu-delta region in a murine hybridoma cell line. The backbone of the gene replacement vector (pCmuCdeltapal) consists of pSV2neo sequences bounded on one side by homology to the mu gene constant (Cmu) region and on the other side by homology to the delta gene constant (Cdelta) region. The Cmu and Cdelta flanking arms of homology were marked by insertions of an identical 30-bp palindrome which frequently escapes mismatch repair when in heteroduplex DNA (hDNA). As a result, intermediates bearing unrepaired hDNA generate mixed (sectored) recombinants following DNA replication and cell division. To monitor the presence and position of sectored sites and, hence, hDNA formation during the recombination process, the palindrome contained a unique NotI site that replaced an endogenous restriction enzyme site at each marker position in the vector-borne Cmu and Cdelta regions. Gene replacement was studied under conditions which permitted the efficient recovery of the product(s) of individual recombination events. Analysis of marker segregation patterns in independent recombinants revealed that extensive hDNA was formed within the Cmu and Cdelta regions. In several recombinants, palindrome markers in the Cmu and Cdelta regions resided on opposite DNA strands (trans configuration). These results are consistent with the mammalian gene replacement reaction involving two crossing-over events in homologous flanking DNA.
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