Open AccessTransient correction of genetic defects in cultured animal cells by introduction of functional proteins.
Author(s) -
Dolores Ortiz,
M M Baldwin,
Joseph J. Lucas
Publication year - 1987
Publication title -
molecular and cellular biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.14
H-Index - 327
eISSN - 1067-8824
pISSN - 0270-7306
DOI - 10.1128/mcb.7.8.3012
Subject(s) - biology , microinjection , cell culture , thymidine kinase , microbiology and biotechnology , thymidine , cell , hypoxanthine guanine phosphoribosyltransferase , guanine , hypoxanthine phosphoribosyltransferase , biochemistry , biophysics , dna , genetics , nucleotide , gene , mutant , virus , herpes simplex virus
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.