Using antisense nucleic acid technology to study the influence of POLκ on genetic stability

AIM:To establish cell line FL-POLκ^- and to study the role ofPOLκ(polymerase kappa) on genetic stability.METHODS:A mammalian expression vector expressing antisense POLκ gene fragment pMAMneo -amp^--POLκ was constructed by cloning the 1 690-1 918 fragment of POLκ gene into the mammalian expression vector pMAMneo-amp^- in antisense orientation. FL cells were fransfected with this antisense RNA expressing vector and selected by G418. Based on the shuttle-plasmid pZ189, the mutation assay was made.RESULTS:The spontaneous mutation frequency of supF tRNA gene in the plasmid replicated in the FL- POLκ^- was 11.2×10^-4, while it was 4.9×10^-4 and 3.7×10^-4 in the control cells FL and FL-M, respectively.CONCLUSION: POLκ playes an important role in maintenance of genetic stability.