Enrichment for gene targeting in mammalian cells by inhibition of poly(ADP-ribosylation)

Waldman, Barbara Criscuolo; O'Quinn, Janis R.; Waldman, Alan S.

doi:10.1016/0167-4781(96)00111-x

Cited by 22 publications

(10 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Cultured mammalian somatic cells are quite proficient at randomly integrating transfected DNA (Waldman et al, 1996), and the frequency of homologous recombination in non-ES cells is typically several orders of magnitude lower than in ES cells (Bollag et al, 1989). The observation that PCR screening failed to detect unambiguous homologous recombination events in more than 100 G418 r -Gc r colonies led to the inference that these colonies might contain randomly integrated homologous recombination vectors, in which the tk gene cassette was destroyed by exonuclease degradation.…”

Section: Discussionmentioning

confidence: 99%

Development of a positive–negative selection procedure for gene targeting in fish cells

2002

View full text Add to dashboard Cite

Section: Discussionmentioning

confidence: 99%

Development of a positive–negative selection procedure for gene targeting in fish cells

2002

View full text Add to dashboard Cite

“…A similar conclusion was drawn from studies in which random integration was significantly inhibited with only marginal effects on homologous recombination. 47,48 Given the involvement of RAD genes in the repair of DSBs by homologous recombination, we were interested to test whether hRAD51 overexpression also led to an enhancement of ionising radiation resistance in human cells. We tested radiation doses between 2.5 and 15 Gy, and found increased survival in hRAD51 overexpressing cells compared with control cells, confirming the involvement of hRAD51 in the repair process.…”

Section: Figure 4 Survival Of Hrad51 Overexpressing Cells To Ionisingmentioning

confidence: 99%

Gene targeting is enhanced in human cells overexpressing hRAD51

Yáñez

Porter

1999

Gene Ther

View full text Add to dashboard Cite

The ideal therapy for single gene disorders would be repair approach, we have overexpressed the gene encoding of the mutated disease genes. Homologous recombination hRAD51, a protein with homologous DNA pairing and is one of several cellular mechanisms for the repair of DNA strand exchange activities, in human cells and measured damage. Recombination between exogenous DNA and its effect on gene targeting. We report a two-to three-fold homologous chromosomal loci (gene targeting) can be increase in gene targeting, and enhanced resistance to used to repair an endogenous gene, but the low efficiency ionising radiation in hRAD51-overexpressing cells with no of this process is a serious barrier to its therapeutic potenobvious detrimental effects. These observations provide tial. Recent progress in the isolation and characterisation valuable genetic evidence for the involvement of hRAD51 of mammalian genes and proteins involved in DNA recomin both gene targeting and DNA repair in human cells. Our bination has raised the possibility that the cellular biochemdata also establish overexpression of recombination genes istry of recombination can be manipulated to improve the as a viable approach to improving gene targeting efficiency of gene targeting. As an initial test of this efficiencies.

show abstract

“…However, the role of this molecule in homologous recombination is not clear. Although chemical inhibition of PARP activity increases chromosomal homologous recombination frequency in mammalian cells (Waldman et al 1996), a transient extrachromosomal recombination study showed that PARP−/− cells can efficiently repair a mutated luciferase reporter gene via homologous recombination (Morrison and Wagner 1996).…”

Section: The Role Of Parp For Other Recombination Events and In Chrommentioning

confidence: 99%

PARP is important for genomic stability but dispensable in apoptosis

Wang¹,

Stingl²,

Morrison³

et al. 1997

Genes Dev.

537

361

View full text Add to dashboard Cite

Mice lacking the gene encoding poly(ADP-ribosyl) transferase (PARP or ADPRT) display no phenotypic abnormalities, although aged mice are susceptible to epidermal hyperplasia and obesity in a mixed genetic background. Whereas embryonic fibroblasts lacking PARP exhibit normal DNA excision repair, they grow more slowly in vitro. Here we investigated the putative roles of PARP in cell proliferation, cell death, radiosensitivity, and DNA recombination, as well as chromosomal stability. We show that the proliferation deficiency in vitro and in vivo is most likely caused by a hypersensitive response to environmental stress. Although PARP is specifically cleaved during apoptosis, cells lacking this molecule apoptosed normally in response to treatment with anti-Fas, tumor neurosis factor ␣, ␥-irradiation, and dexamethasone, indicating that PARP is dispensable in apoptosis and that PARP−/− thymocytes are not hypersensitive to ionizing radiation. Furthermore, the capacity of mutant cells to carry out immunoglobulin class switching and V(D)J recombination is normal. Finally, primary PARP mutant fibroblasts and splenocytes exhibited an elevated frequency of spontaneous sister chromatid exchanges and elevated micronuclei formation after treatment with genotoxic agents, establishing an important role for PARP in the maintenance of genomic integrity.

show abstract

Enrichment for gene targeting in mammalian cells by inhibition of poly(ADP-ribosylation)

Cited by 22 publications

References 16 publications

Development of a positive–negative selection procedure for gene targeting in fish cells

Development of a positive–negative selection procedure for gene targeting in fish cells

Gene targeting is enhanced in human cells overexpressing hRAD51

PARP is important for genomic stability but dispensable in apoptosis

Contact Info

Product

Resources

About