Insertion Mutagenesis of Embryonal Carcinoma Cells by Retroviruses

King, Walter E.; Patel, M. S.; Lobel, L I; Sp, Goff; Nguyen-Huu, M C

doi:10.1126/science.3838595

Cited by 126 publications

(61 citation statements)

References 25 publications

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“…But this is also proportional to the higher risk of mutagenic events. Previous studies on retroviral-induced mutagenesis in mammalian cells have found a ratio of "mutations versus insertional events" which ranged from 10 -9 to 10 -3 [184][185][186][187][188][189]. Such variability indicates that the ratio of mutations per insertional events depends on the cell type and assay system.…”

Section: Vector Systems Based On Retroviruses Lentiviruses and Foamimentioning

confidence: 99%

Latest Developments in Gene Transfer Technology: Achievements, Perspectives, and Controversies over Therapeutic Applications

et al. 2000

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Section: Vector Systems Based On Retroviruses Lentiviruses and Foamimentioning

confidence: 99%

Latest Developments in Gene Transfer Technology: Achievements, Perspectives, and Controversies over Therapeutic Applications

et al. 2000

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“…The possible cause of the relatively small increase in hprt mutagenesis to 7.4 Â 10 À6 (2.3-fold above the spontaneous mutation frequency) observed following the cocultivation infection procedure became more apparent after Southern analysis revealed between 1 and 19 provirus insertion events and rearrangements in the hprt gene in 63% of the mutants examined. Chromosomal rearrangements have also been observed using replication competent retroviruses by King et al 19 Between 5 and 50 provirus insertions were identified per mutant and the frequency of mutagenesis was 10…”

Section: Retrieval Of the Lhl Provirus From The Hprt Locusmentioning

confidence: 99%

“…In practice, however, using replication-competent viruses only one in 10 8 provirus insertions caused mutagenesis in this region. 19 Evaluating the risk of insertional mutagenesis using immortal cell lines must be treated with caution, however. For example several immortal cell lines have been shown to carry mutant p53, which has been implicated in many chromosomal abberations in tumours.…”

Section: Retrieval Of the Lhl Provirus From The Hprt Locusmentioning

confidence: 99%

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Mutational effects of retrovirus insertion on the genome of V79 cells by an attenuated retrovirus vector: implications for gene therapy

et al. 2003

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Attenuated retroviruses are currently the most widely used vectors in clinical gene therapy because of their potential to effect stable and permanent gene transfer. Since gene delivery is accompanied by random insertion of foreign genetic material into the recipient chromosomal DNA, the potential for insertional mutagenesis exists. In this study, we used a defective retrovirus vector containing a selectable marker, the hygromycin phosphotransferase gene, to investigate the mutagenic effects of vector integration on the mammalian genome. V79 Chinese hamster cells were infected with virus supernatants or by coculture with virus producer cells, and provirus insertion events occurred at low and high frequencies, respectively. The frequency of hprt mutagenesis was increased by a factor of 2.3 over the spontaneous hprt mutation frequency only following multiple provirus insertions/cell genome. Multiple provirus insertions (43/genome) resulted in instability at the hprt locus in 63% of the virally induced hprt mutants, as indicated by rearrangements at the molecular level, whereas no rearrangements were found when the provirus copy number was 1-2/genome. To demonstrate direct proviral involvement in mutagenesis, the defective MLV vector was retrieved along with flanking genomic hprt sequences from one mutant, and localized within intron 5 of the hprt gene. These data suggest that provirus copy number is a key factor when considering the potential hazards of using retrovirus vectors for gene therapy.

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“…This observation suggested that the proviral genome (endogenous ecotropic murine leukemia viral locus 15, Emv-15) is physically close to or within the agouti locus and that isolation of the provirus would allow molecular entry to be gained into the region of the agouti locus on chromosome 2. The fact that proviral integration can result in activation (9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20)(21) or inactivation (22)(23)(24)(25)(26)(27) MATERIALS AND METHODS Mice. All animals were obtained from The Jackson Laboratory.…”

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confidence: 99%

The lethal yellow allele-associated provirus results in the production of chimeric viral-host RNAs.

Lovett¹,

Epstein²

1987

Proc. Natl. Acad. Sci. U.S.A.

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The lethal yellow (A') mutation is the most dominant allele at the agouti locus of the mouse. Mice heterozygous for this allele have a yellow coat color, are genetically obese with an increased susceptibility to cancer, and have other metabolic derangements. Mice homozygous for Ay die early in embryogenesis, possibly because of a trophectoderm defect. The Ay mutation is distinguished from the many alleles described at the agouti locus in being associated with an endogenous provirus, designated endogenous ecotropic murine leukemia viral locus 15 (Emv-15). To obtain DNA sequences from regions close to or within the agouti locus, we have isolated the Emv-15 provirus and have found that the DNA sequences adjacent to the provirus are part of a mouse gene that is expressed in the same transcriptional orientation as the proviral genes. These mouse DNA sequences recognize two distinct size classes of RNA in various adult wild-type tissues including skin. In Ay heterozygotes both types of transcript hybridize to the proviral long terminal repeat, and, in heterozygous spleens, the shorter transcript is present at an enhanced steady-state level. These results suggest that the Ayassociated provirus (Emv-15) is distinguished from the endogenous C-type proviruses described to date in having altered the expression of a flanking host gene by promoter insertion, resulting in the production of chimeric viral-host fusion transcripts.The agouti locus (a) is a coat color-determining locus in the mouse that is located on chromosome 2 and that controls the relative amount and distribution of yellow (phaeomelanin) and black/brown (eumelanin) pigment in the hairs of the coat (1, 2).

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Insertion Mutagenesis of Embryonal Carcinoma Cells by Retroviruses

Cited by 126 publications

References 25 publications

Latest Developments in Gene Transfer Technology: Achievements, Perspectives, and Controversies over Therapeutic Applications

Latest Developments in Gene Transfer Technology: Achievements, Perspectives, and Controversies over Therapeutic Applications

Mutational effects of retrovirus insertion on the genome of V79 cells by an attenuated retrovirus vector: implications for gene therapy

The lethal yellow allele-associated provirus results in the production of chimeric viral-host RNAs.

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