Gene Targeting in Rat Embryo Fibroblasts Promoted by the Polyomavirus Large T Antigen

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1996 Jun 1

PMID 8668528

Authors

V Frances

M Bastin

Affiliations

Soon will be listed here.

Abstract

We used the recombination-promoting activity of the polyomavirus large T antigen (T-ag) to increase the frequency of gene targeting in rat fibroblasts. We constructed a cell line carrying a functional polyomavirus replication origin and a transformation-defective middle T-ag oncogene. The structure of the locus was such that homologous recombination with the targeting DNA reconstituted a functional transforming gene and converted the cells from the normal to the transformed state. Introduction of the large T-ag with the targeting DNA promoted recombinational events that corrected the mutation in either the target locus or the targeting DNA. The frequency of recombination was not substantially influenced by the extent of homology between the recombining sequences. However, it was reduced when the replication origin was inactivated in the targeting DNA, and was reduced further when the origin was inactivated in the target locus.

References

Joyner A, Herrup K, Auerbach B, Davis C, Rossant J . Subtle cerebellar phenotype in mice homozygous for a targeted deletion of the En-2 homeobox. Science. 1991; 251(4998):1239-43. DOI: 10.1126/science.1672471. View

Laurent S, Frances V, Bastin M . Intrachromosomal recombination mediated by the polyomavirus large T antigen. Virology. 1995; 206(1):227-33. DOI: 10.1016/s0042-6822(95)80037-9. View

Accili D, Taylor S . Targeted inactivation of the insulin receptor gene in mouse 3T3-L1 fibroblasts via homologous recombination. Proc Natl Acad Sci U S A. 1991; 88(11):4708-12. PMC: 51735. DOI: 10.1073/pnas.88.11.4708. View

Valancius V, SMITHIES O . Double-strand gap repair in a mammalian gene targeting reaction. Mol Cell Biol. 1991; 11(9):4389-97. PMC: 361301. DOI: 10.1128/mcb.11.9.4389-4397.1991. View

Hasty P, Chang C, Bradley A . Target frequency and integration pattern for insertion and replacement vectors in embryonic stem cells. Mol Cell Biol. 1991; 11(9):4509-17. PMC: 361323. DOI: 10.1128/mcb.11.9.4509-4517.1991. View

St-Onge L, Bastin M . Amplification of polyomavirus DNA sequences stably integrated in rat cells. Nucleic Acids Res. 1991; 19(23):6619-25. PMC: 329231. DOI: 10.1093/nar/19.23.6619. View

Kim C, Epner E, Forrester W, Groudine M . Inactivation of the human beta-globin gene by targeted insertion into the beta-globin locus control region. Genes Dev. 1992; 6(6):928-38. DOI: 10.1101/gad.6.6.928. View

Te Riele H, Maandag E, Berns A . Highly efficient gene targeting in embryonic stem cells through homologous recombination with isogenic DNA constructs. Proc Natl Acad Sci U S A. 1992; 89(11):5128-32. PMC: 49242. DOI: 10.1073/pnas.89.11.5128. View

Botchan M, Topp W, Sambrook J . Studies on simian virus 40 excision from cellular chromosomes. Cold Spring Harb Symp Quant Biol. 1979; 43 Pt 2:709-19. DOI: 10.1101/sqb.1979.043.01.079. View

10.

Colantuoni V, Dailey L, Valle G, Basilico C . Requirements for excision and amplification of integrated viral DNA molecules in polyoma virus-transformed cells. J Virol. 1982; 43(2):617-28. PMC: 256164. DOI: 10.1128/JVI.43.2.617-628.1982. View

11.

Watanabe S, Temin H . Construction of a helper cell line for avian reticuloendotheliosis virus cloning vectors. Mol Cell Biol. 1983; 3(12):2241-9. PMC: 370095. DOI: 10.1128/mcb.3.12.2241-2249.1983. View

12.

Miller J, Bullock P, Botchan M . Simian virus 40 T antigen is required for viral excision from chromosomes. Proc Natl Acad Sci U S A. 1984; 81(23):7534-8. PMC: 392181. DOI: 10.1073/pnas.81.23.7534. View

13.

Rubnitz J, Subramani S . The minimum amount of homology required for homologous recombination in mammalian cells. Mol Cell Biol. 1984; 4(11):2253-8. PMC: 369052. DOI: 10.1128/mcb.4.11.2253-2258.1984. View

14.

Subramani S, Rubnitz J . Recombination events after transient infection and stable integration of DNA into mouse cells. Mol Cell Biol. 1985; 5(4):659-66. PMC: 366767. DOI: 10.1128/mcb.5.4.659-666.1985. View

15.

Jasin M, De Villiers J, Weber F, Schaffner W . High frequency of homologous recombination in mammalian cells between endogenous and introduced SV40 genomes. Cell. 1985; 43(3 Pt 2):695-703. DOI: 10.1016/0092-8674(85)90242-9. View

16.

Wong E, Capecchi M . Analysis of homologous recombination in cultured mammalian cells in transient expression and stable transformation assays. Somat Cell Mol Genet. 1986; 12(1):63-72. DOI: 10.1007/BF01560728. View

17.

Ayares D, Chekuri L, Song K, Kucherlapati R . Sequence homology requirements for intermolecular recombination in mammalian cells. Proc Natl Acad Sci U S A. 1986; 83(14):5199-203. PMC: 323918. DOI: 10.1073/pnas.83.14.5199. View

18.

Rauth S, Song K, Ayares D, Wallace L, Moore P, Kucherlapati R . Transfection and homologous recombination involving single-stranded DNA substrates in mammalian cells and nuclear extracts. Proc Natl Acad Sci U S A. 1986; 83(15):5587-91. PMC: 386333. DOI: 10.1073/pnas.83.15.5587. View

19.

Song K, Chekuri L, Rauth S, Ehrlich S, Kucherlapati R . Effect of double-strand breaks on homologous recombination in mammalian cells and extracts. Mol Cell Biol. 1985; 5(12):3331-6. PMC: 369160. DOI: 10.1128/mcb.5.12.3331-3336.1985. View

20.

Wake C, Vernaleone F, Wilson J . Topological requirements for homologous recombination among DNA molecules transfected into mammalian cells. Mol Cell Biol. 1985; 5(8):2080-9. PMC: 366926. DOI: 10.1128/mcb.5.8.2080-2089.1985. View