Construction of HPV16 E6 siRNA expressing recombinants and identification of transfection efficiency in vitro / 重庆医科大学学报

ABSTRACT

Objective:To silence the expression of HPV16 E6 gene in Ca Ski cell line with RNA interference technique.In order to understand if the HPV16 E6 siRNA could inhibit the expression of E6 gene specifically,recombinant eukaryotic vectors were constructed and transfected into Ca Ski cell line.The transfection efficiency was observed.Methods: According to the sequence of HPV16 E6 gene,the E6 siRNA was designed and cloned into the EGFP reporter plasmid pGenesil-1,and then transfected into CaSki cell line.The fluorescence expression was observed and the transfection efficiency was identified.Results: Two siRNA expressing recombinants and the corresponding negative control vector were constructed and transfected into Ca Ski cell successfully,and the transfection efficiency achieved about 50%.Conclusion: The construction of recombinant E6 siRNA eukaryotic vectors and transfection of them into Ca Ski cell line successfully established a favourable foundation for further study.