DNA damaging agents improve stable gene transfer efficiency in mammalian cells

Stevens, Craig W.; Cerniglia, George J.; Giandomenico, Albert R.; Koch, Cameron J.

doi:10.1002/(sici)1520-6823(1998)6:1<1::aid-roi1>3.0.co;2-1

Cited by 9 publications

(6 citation statements)

References 41 publications

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“…Previous studies have shown that ionizing radiation enhances the frequency of nonhomologous integration in yeast and mammalian cells (33, 34). In addition, other damaging agents, including camptothecin, VP16, UV radiation and hydrogen peroxide, increase nonhomologous integration of transfected plasmids in mammalian cells (35-37). However, extensive sequence analysis of DNA damage-induced illegitimate recombination events has yet to be carried out.…”

Section: Introductionmentioning

confidence: 99%

Ionizing Radiation Induces Microhomology-Mediated End Joiningin transin Yeast and Mammalian Cells

Scuric¹,

Chan

Hafer

et al. 2009

Radiation Research

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DNA double-strand breaks repaired through nonhomologous end joining require no extended sequence homology as a template for the repair. A subset of end-joining events, termed microhomology-mediated end joining, occur between a few base pairs of homology, and such pathways have been implicated in different human cancers and genetic diseases. Here we investigated the effect of exposure of yeast and mammalian cells to ionizing radiation on the frequency and mechanism of rejoining of transfected unirradiated linear plasmid DNA. Cells were exposed to γ radiation prior to plasmid transfection; subsequently the rejoined plasmids were recovered and the junction sequences were analyzed. In irradiated yeast cells, 68% of recovered plasmids contained microhomologies, compared to only 30% from unirradiated cells. Among them 57% of events used ≥4 bp of microhomology compared to only 11% from unirradiated cells. In irradiated mammalian cells, 54% of plasmids used ≥4 bp of microhomology compared to none from unirradiated cells. We conclude that exposure of yeast and mammalian cells to radiation prior to plasmid transfection enhances the frequency of microhomology-mediated end-joining events in trans. If such events occur within genomic locations, they may be involved in the generation of large deletions and other chromosomal aberrations that occur in cancer cells.

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Section: Introductionmentioning

confidence: 99%

Ionizing Radiation Induces Microhomology-Mediated End Joiningin transin Yeast and Mammalian Cells

Scuric¹,

Chan

Hafer

et al. 2009

Radiation Research

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“…Ionizing radiation enhances the frequency of nonhomologous integration (NHI) in yeast ( 19 ). In addition, other DNA damaging agents including UV, bleomycin, camptothecin, VP-16 and hydrogen peroxide also enhance gene integration in mammalian cells ( 20–25 ). These DNA damaging agents may eventually lead to the formation of DSBs and suggest that DSBs could be the inducing agent for the enhancement of integration.…”

Section: Introductionmentioning

confidence: 99%

Ionizing radiation and restriction enzymes induce microhomology-mediated illegitimate recombination in Saccharomyces cerevisiae

Chan¹,

Kiechle²,

Manivasakam³

et al. 2007

Nucleic Acids Research

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DNA double-strand breaks can be repaired by illegitimate recombination without extended sequence homology. A distinct mechanism namely microhomology-mediated recombination occurs between a few basepairs of homology that is associated with deletions. Ionizing radiation and restriction enzymes have been shown to increase the frequency of nonhomologous integration in yeast. However, the mechanism of such enhanced recombination events is not known. Here, we report that both ionizing radiation and restriction enzymes increase the frequency of microhomology-mediated integration. Irradiated yeast cells displayed 77% microhomology-mediated integration, compared to 27% in unirradiated cells. Radiation-induced integration exhibited lack of deletions at genomic insertion sites, implying that such events are likely to occur at undamaged sites. Restriction enzymes also enhanced integration events at random non-restriction sites via microhomology-mediated recombination. Furthermore, generation of a site-specific I-SceI-mediated double-strand break induces microhomology-mediated integration randomly throughout the genome. Taken together, these results suggest that double-strand breaks induce a genome-wide microhomology-mediated illegitimate recombination pathway that facilitates integration probably in trans at non-targeted sites and might be involved in generation of large deletions and other genomic rearrangements.

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“…REI requires that the transfected plasmid be damaged (containing either double-or single-strand DNA breaks (Stevens et al 1996(Stevens et al , 1998, but such damage does not affect transfection efficiency in unirradiated cells. Ku80 is required for plasmid integration in both irradiated and unirradiated cells (Stevens et al 1999).…”

Section: Discussionmentioning

confidence: 99%

“…X-rays (Stevens et al 1996), ultraviolet (UV) light (Perez et al 1985), H 2 O 2 (Stevens et al 1998), psoralen crosslinking (Vos and Hanawalt 1989), cisplatin (Son and Huang 1994) and electroporation with restriction endonucleases (Puchta et al 1993) all enhance recombination in mammalian cells, as measured by the degree of increased plasmid integration. Radiation improves the integration of linearized, but not supercoiled, plasmids (Zeng et al 1997).…”

Section: Introductionmentioning

confidence: 99%

Nucleotide excision repair proteins and their importance for radiation‐enhanced transfection

Nimura¹,

Prithivirajsingh

Ismail

et al. 2003

International Journal of Radiation Biology

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DNA damaging agents improve stable gene transfer efficiency in mammalian cells

Cited by 9 publications

References 41 publications

Ionizing Radiation Induces Microhomology-Mediated End Joiningin transin Yeast and Mammalian Cells

Ionizing Radiation Induces Microhomology-Mediated End Joiningin transin Yeast and Mammalian Cells

Ionizing radiation and restriction enzymes induce microhomology-mediated illegitimate recombination in Saccharomyces cerevisiae

Nucleotide excision repair proteins and their importance for radiation‐enhanced transfection

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