Differences in the efficiency and stability of gene expression after transfection and nuclear injection: a study with a chick delta-crystallin gene.
The efficiency of an exogenous gene's expression was compared after its transfection and injection into various mouse cells to systematically evaluate these two gene transfer techniques. Special attention was paid to the period of transient expression. The gene used was a derivative of chicken delta-crystallin gene with the 5' end region replaced by a promoter base sequence of a retrovirus. Nuclear injection was more efficient than transfection in several respects: it was roughly one thousand times more efficient in producing gene-expressing cells than the transfection technique; it produced positive cells in every challenged cell line in contrast to the results of some unsuccessful trials found with transfection; and the maximum expression of the exogenous gene in a gene-transferred cell was much higher after injection than after transfection. With the transfection technique, use of a DNA-calcium phosphate coprecipitate was slightly more efficient than the use of DEAE-dextran. The stability of gene expression in transfected and nuclear-injected cells differed greatly: Expression of the exogenous gene in transfected cells was transmitted to 92% of the daughter cells per division, whereas its expression in injected cells was transmitted to only 32% of the daughter cells. This great difference in stability probably reflects different states of the major fraction of the exogenous gene: integration into chromosomes in transfected cells versus extrachromosomal localization in injected cells.
D000818:Animals; D002460:Cell Line; D002645:Chickens / Q000235:genetics; D003459:Crystallins / Q000096:biosynthesis / Q000235:genetics*; D004274:DNA, Recombinant; D005786:Gene Expression Regulation; D051379:Mice; D008845:Microinjections; D009876:Operon; D014162:Transfection; D014170:Transformation, Genetic
D003459:Crystallins; D004274:DNA, Recombinant