ArticleChromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology2010
Integration of exogenous DNA into mouse embryonic stem cell chromosomes shows preference into genes and frequent modification at junctions.
Article in Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.
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4 citing papers in PubMed, 5 citations in OpenAlex.
- Efficient multi-kilobase knock-ins in mice and cell lines using CRISPR/Cas9 and rAAV donors with unbiased whole-genome characterization by LOCK-seq.Nucleic acids research · 2026Article
- Dual loss of human POLQ and LIG4 abolishes random integration.Nature communications · 2017Article
- Construction and applications of exon-trapping gene-targeting vectors with a novel strategy for negative selection.BMC research notes · 2015Article
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7 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Chromosomal integration of exogenous DNA in mammalian cells allows stable gene expression for a variety of biological applications. Although it is presumably mediated by DNA repair machinery, little is known regarding site preferences and other characteristics. We isolated and analyzed 256 chromosomal-plasmid DNA integration junctions from 158 plasmid integrants after electroporation in mouse embryonic stem (ES) cells. The frequency of integrations in transcription units (40%) showed a slight but significant increase over the frequency estimated by computer simulation of random events (30%), suggesting preferential integration into genes. Microarray analysis revealed preference into genes, which are expressed in mouse ES cells. In contrast, bias toward integrations around transcriptional start sites, CpG islands and repeat elements was not observed. Furthermore, all host chromosome sequences as well as the majority of plasmids (96%) at the integration junctions were modified by deletions and/or insertions of additional nucleotides. Detailed analyses revealed frequent stem loop/hairpin formation mediated by weak homologies near plasmid ends before integration. Our study sheds light on a natural fate of exogenous DNA, which preferentially integrates into transcriptionally active chromosomal sites and by an imprecise end-joining pathway, associated with highly frequent modification of the end sequences.
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Registered trials
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