Mouse transgenes in human cells detect specific base substitutions

D. A. Schaff, R. A. Jarrett, Stephen Dlouhy, S. Ponniah, M. Stockelman, P. J. Stambrook, J. A. Tischfield

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

We describe a system of transgenic human cell lines that detects and identifies specific point mutations at defined positions within a gene. The target transgenome is a mouse adenine phosphoribosyltransferase (APRT) gene rendered nonfunctional by introduction of a substitution at either of two bases that comprise a splice acceptor site. Reversion at a mutated site results in the expression of wild-type mouse APRT and consequent growth of APRT+ transgenic cell colonies. Site-specific reversion to wild-type sequence is confirmed by regeneration of a previously destroyed diagnostic Pst I site. Two independent cell clones, each with mutant transgenomes bearing an A → G transition, exhibited an up to 7500-fold, dose-dependent induction of reversion following treatment with ethyl methanesulfonate. Treatment of these clones with 2-aminopurine resulted in no induction of revertants. In contrast, another transgenic cell clone, bearing a G → A transition, reverted as a consequence of 2-aminopurine, but not ethyl methanesulfonate, treatment. These data confirm for human cells the proposed mechanisms of action of these mutagens and provide evidence for the utility of our site-specific reversion method for mutagenesis studies.

Original languageEnglish
Pages (from-to)8675-8679
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume87
Issue number21
StatePublished - 1990

Fingerprint

Adenine Phosphoribosyltransferase
2-Aminopurine
Transgenes
Ethyl Methanesulfonate
Clone Cells
RNA Splice Sites
Mutagens
Point Mutation
Mutagenesis
Genes
Regeneration
Cell Line
Growth

Keywords

  • 2-aminopurine
  • Adenine phosphoribosyltransferase
  • Ethyl methanesulfonate
  • Mutagenesis
  • Splice junction

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Schaff, D. A., Jarrett, R. A., Dlouhy, S., Ponniah, S., Stockelman, M., Stambrook, P. J., & Tischfield, J. A. (1990). Mouse transgenes in human cells detect specific base substitutions. Proceedings of the National Academy of Sciences of the United States of America, 87(21), 8675-8679.

Mouse transgenes in human cells detect specific base substitutions. / Schaff, D. A.; Jarrett, R. A.; Dlouhy, Stephen; Ponniah, S.; Stockelman, M.; Stambrook, P. J.; Tischfield, J. A.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 87, No. 21, 1990, p. 8675-8679.

Research output: Contribution to journalArticle

Schaff, DA, Jarrett, RA, Dlouhy, S, Ponniah, S, Stockelman, M, Stambrook, PJ & Tischfield, JA 1990, 'Mouse transgenes in human cells detect specific base substitutions', Proceedings of the National Academy of Sciences of the United States of America, vol. 87, no. 21, pp. 8675-8679.
Schaff, D. A. ; Jarrett, R. A. ; Dlouhy, Stephen ; Ponniah, S. ; Stockelman, M. ; Stambrook, P. J. ; Tischfield, J. A. / Mouse transgenes in human cells detect specific base substitutions. In: Proceedings of the National Academy of Sciences of the United States of America. 1990 ; Vol. 87, No. 21. pp. 8675-8679.
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