Skip to main content
NCBI home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1987 Aug;7(8):3012–3017. doi: 10.1128/mcb.7.8.3012

Transient correction of genetic defects in cultured animal cells by introduction of functional proteins.

D Ortiz 1, M M Baldwin 1, J J Lucas 1
PMCID: PMC367926  PMID: 3670304

Abstract

Material was introduced into cultures of cells by using the method of scrape loading, in which cells are simply rubbed from the surface of a plastic tissue culture dish by a rubber-tipped rod in the presence of a macromolecule of interest. The volume of solution introduced into cells was comparable to that generally injected in the direct microinjection method with glass capillaries, that is, about 50 to 100 fl per cell. Genetic defects (lack of hypoxanthine-guanine phosphoribosyltransferase and thymidine kinase) in several cell lines were transiently corrected by scraping the cells in the presence of crude cell extracts prepared from wild-type cells.

Full text

PDF
3012

Images in this article

3013Image
on p.3013
3015Image
on p.3015
3016Image
on p.3016

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bar-Sagi D., Feramisco J. R. Induction of membrane ruffling and fluid-phase pinocytosis in quiescent fibroblasts by ras proteins. Science. 1986 Sep 5;233(4768):1061–1068. doi: 10.1126/science.3090687. [DOI] [PubMed] [Google Scholar]
  2. Celis J. E. Microinjection of somatic cells with micropipettes: comparison with other transfer techniques. Biochem J. 1984 Oct 15;223(2):281–291. doi: 10.1042/bj2230281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Croce C. M., Girardi A. J., Koprowski H. Assignment of the T-antigen gene of simian virus 40 to human chromosome C-7. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3617–3620. doi: 10.1073/pnas.70.12.3617. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Graessmann M., Graessmann A. Microinjection of tissue culture cells. Methods Enzymol. 1983;101:482–492. doi: 10.1016/0076-6879(83)01033-2. [DOI] [PubMed] [Google Scholar]
  5. Hightower M. J., Lucas J. J. Construction of viable mouse-human hybrid cells by nuclear transplantation. J Cell Physiol. 1980 Oct;105(1):93–103. doi: 10.1002/jcp.1041050112. [DOI] [PubMed] [Google Scholar]
  6. LITTLEFIELD J. W. THREE DEGREES OF GUANYLIC ACID--INOSINIC ACID PYROPHOSPHORYLASE DEFICIENCY IN MOUSE FIBROBLASTS. Nature. 1964 Sep 12;203:1142–1144. doi: 10.1038/2031142a0. [DOI] [PubMed] [Google Scholar]
  7. Lipsich L. A., Lucas J. J., Kates J. R. Cell cycle dependence of the reactivation of chick erythrocyte nuclei after transplantation into mouse L929 cell cytoplasts. J Cell Physiol. 1978 Nov;97(2):199–207. doi: 10.1002/jcp.1040970209. [DOI] [PubMed] [Google Scholar]
  8. Lucas J. J., Kates J. R. Nuclear transplantation with mammalian cells. Methods Cell Biol. 1977;15:359–370. doi: 10.1016/s0091-679x(08)60225-3. [DOI] [PubMed] [Google Scholar]
  9. McNeil P. L., Murphy R. F., Lanni F., Taylor D. L. A method for incorporating macromolecules into adherent cells. J Cell Biol. 1984 Apr;98(4):1556–1564. doi: 10.1083/jcb.98.4.1556. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Thompson E. B., Tomkins G. M., Curran J. F. Induction of tyrosine alpha-ketoglutarate transaminase by steroid hormones in a newly established tissue culture cell line. Proc Natl Acad Sci U S A. 1966 Jul;56(1):296–303. doi: 10.1073/pnas.56.1.296. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Wang Y. L. Exchange of actin subunits at the leading edge of living fibroblasts: possible role of treadmilling. J Cell Biol. 1985 Aug;101(2):597–602. doi: 10.1083/jcb.101.2.597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. White J. D., Bruno J., Lucas J. J. Programmed macromolecular synthesis in regenerating karyoplasts. Mol Cell Biol. 1983 Oct;3(10):1866–1881. doi: 10.1128/mcb.3.10.1866. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wigler M., Pellicer A., Silverstein S., Axel R., Urlaub G., Chasin L. DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1373–1376. doi: 10.1073/pnas.76.3.1373. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES