Skip to main content
Log in

Production of transmitochondrial mouse cell lines by cybrid rescue of rhodamine-6G pre-treated L-cells

  • Brief Communication
  • Published:
Somatic Cell and Molecular Genetics

Abstract

Treatment of mouse LMTK cells with the toxic mitochondrial dye rhodamine 6G (R-6G) at 2.5 μg/ml for 7 days prevented cell growth while maintaining viability, with less than 10−6 cells recovering to form colonies. Pre-treatment of LMTK cells with R-6G was followed by fusion with enucleated mouse 501–1 cells harboring a homoplasmic point mutation in the mitochondrial DNA (mtDNA) 16S rRNA gene conferring chloramphenicol resistance (CAPR). Cybrids and any surviving unfused LMTK cells were selected in BrdU with or without CAP and their mtDNAs screened for the presence of the CAPR marker. Approximately 1 colony per 2×105 LMTK cells appeared in the fusion plates selected both with and without CAP. Most clones investigated were confirmed to be cybrids by showing the presence of the generally homoplasmic CAPR mutation, whether or not CAP selection was used. Hence, R-6G pre-treatment permits construction of transmitochondrial cybrid cell lines carrying a variety of mtDNAs, without the need for ρo cell lines.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Literature Cited

  1. Morais, R., Desjardins, P., Turmel, C., and Zinkewich-Peotti, K. (1987).In Vitro Cell Devel. Biol. 24:649–658.

    Google Scholar 

  2. King, M., and Attardi, G. (1989).Science 246:500–503.

    CAS  PubMed  Google Scholar 

  3. Wallace, D.C., Bunn, C.L., and Eisenstadt, J.M. (1975).J. Cell Biol. 67:174–188.

    Article  CAS  PubMed  Google Scholar 

  4. Chomyn, A., Meola, G., Bresolin, N., Lai, S.T., Scarlato, G., and Attardi, G. (1991).Mol. Cell. Biol. 11:2236–2244.

    CAS  PubMed  Google Scholar 

  5. Hayashi, J-I., Ohta, S., Kikuchi, A., Takemitsu, M., Goto, Y-I., and Nonaka, I. (1991).Proc. Natl. Acad. Sci. U.S.A. 88:10614–10618.

    CAS  PubMed  Google Scholar 

  6. King, M., Koga, Y., Davidson, M., and Schon, E.A. (1992).Mol. Cell. Biol. 12:480–490.

    CAS  PubMed  Google Scholar 

  7. Trounce, I., Neill, S., and Wallace, D.C. (1994)Proc. Natl. Acad. Sci. U.S.A. 91:8334–8338.

    CAS  PubMed  Google Scholar 

  8. Ziegler, M.L., and Davidson, R.L. (1981).Somatic Cell Genet. 7:73–88.

    Article  CAS  PubMed  Google Scholar 

  9. Lampidis, T.J., Hasin, Y., Weiss, M.J., and Chen, L.B. (1985).Biomed. Pharmacother. 39:220–226.

    CAS  PubMed  Google Scholar 

  10. Chen, L.B. (1988).Ann. Rev. Cell Biol. 4:155–181.

    CAS  PubMed  Google Scholar 

  11. Mannella, C.A., and Wang, Q. (1989).Biochem. Biophys. Acta 981:363–366.

    CAS  PubMed  Google Scholar 

  12. Johnson, L.V., Walsh, M.L., and Chen, L.B. (1980).Proc. Natl. Acad. Sci. U.S.A. 77:990–994.

    CAS  PubMed  Google Scholar 

  13. Gear, A.R.L. (1974).J. Biol. Chem. 249:3628–3637.

    CAS  PubMed  Google Scholar 

  14. Higuti, T., Niimi, S., Saito, R., Nakasima, S.N., Ohe, T., Tani, I., and Yoshimura, T. (1980).Biochem. Biophys. Acta 593:463–467.

    CAS  PubMed  Google Scholar 

  15. Mai, M.S., and Allison, W.S. (1983).Arch. Biochem. Biophys. 221:467–476.

    Article  CAS  PubMed  Google Scholar 

  16. Wieker, H-J., Kuschmitz, D., and Hess, B. (1987).Biochem. Biophys. Acta 892:108–117.

    CAS  PubMed  Google Scholar 

  17. Bullough, D.A., Ceccarelli, E.A., Roise, D., and Allison, W.S. (1989).Biochem. Biophys. Acta 975:377–383.

    CAS  PubMed  Google Scholar 

  18. Kit, S., Dubbs, D.R., Piekarski, L.J., and Hsu, T.C. (1963).Exp. Cell. Res. 31:297–312.

    CAS  PubMed  Google Scholar 

  19. Littlefield, J.W. (1964).Nature 203:1142–1144.

    CAS  PubMed  Google Scholar 

  20. Bunn, C.L., Wallace, D.C., and Eisenstadt, J.M. (1974).Proc. Natl. Acad. Sci. U.S.A. 71:1681–1685.

    CAS  PubMed  Google Scholar 

  21. Bunn, C.L., Wallace, D.C., and Eisenstadt, J.M. (1977).Somatic Cell Molec. Genet. 3:71–92.

    CAS  Google Scholar 

  22. Blanc, H., Wright, C.T., Bibb, M.J., Wallace, D.C., and Clayton, D.A. (1981).Proc. Natl. Acad. Sci. U.S.A. 78:3789–3793.

    CAS  PubMed  Google Scholar 

  23. Bibb, M.J., Van Etten, R.A., Wright, C.T., Walberg, M.W., and Clayton, D.A. (1981).Cell 26:167–180.

    Article  CAS  PubMed  Google Scholar 

  24. Sandford, J.A., and Stubblefield, E. (1987).Somatic Cell Molec. Genet. 13:279–284.

    Google Scholar 

  25. Ziegler, M.L. (1982). InTechniques in Somatic Cell Genetics, (ed.) Shay, J.W. Plenum, New York, pp. 211–219.

    Google Scholar 

  26. Smith, R., Huston, M.M., Jenkins, R.N., Huston, D.P., and Rich, R.R. (1983).Nature 306:599–601.

    CAS  PubMed  Google Scholar 

  27. Zinkewich-Peotti, K., Parent, M., and Morais, R. (1990).Cancer Res. 50:6675–6682.

    CAS  PubMed  Google Scholar 

  28. Terasaki, M., Song, J., Wong, J.R., Weiss, M.J., and Chen, L.B. (1984).Cell 38:101–108.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Trounce, I., Wallace, D.C. Production of transmitochondrial mouse cell lines by cybrid rescue of rhodamine-6G pre-treated L-cells. Somat Cell Mol Genet 22, 81–85 (1996). https://doi.org/10.1007/BF02374379

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02374379

Keywords

Navigation