Analysis of electroporation-induced genetic damages in V79/AP4 Chinese hamster cells.
Electroporation is a recent technique used to introduce exogenous DNA into eukaryotic cells. It is important to establish that the gene of interest is transferred into a functional, non-mutated recipient cell. V79/AP4 Chinese hamster cells were exposed to high-voltage pulsed electric fields and some biological and genetic effects were measured. The results showed that cytotoxicity was related in a dose-dependent manner to the number of applied pulses. Thioguanine-resistant colony-forming cells as well as chromosomal aberrations were also induced whereas ouabain resistants and sister-chromatid exchanges were not or slightly induced. Spontaneous and electroporation-induced clones that were phenotypically TGR/HATS were used to investigate the hprt locus. Molecular screening of the locus showed that the number of deleted exons was significantly higher in induced than in spontaneous TG-resistant clones, suggesting that the genetic damages induced by electroporation concern the loss of regions well over the size of the hprt locus.
Vatteroni
L
L
Istituto di Mutagenesi e Differenziamento, CNR, Pisa, Italy.
Piras
A
A
Simi
S
S
Mariani
L
L
Moretti
A
A
Citti
L
L
Mariani
T
T
Rainaldi
G
G
D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D002869:Chromosome Aberrations; D006224:Cricetinae; D003412:Cricetulus; D004249:DNA Damage; D004560:Electricity; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D009154:Mutation; D016133:Polymerase Chain Reaction; D012854:Sister Chromatid Exchange; D013866:Thioguanine / Q000494:pharmacology
D007041:Hypoxanthine Phosphoribosyltransferase; D013866:Thioguanine