Construction and effect identification of RNAi eukaryotic expression vectors of iASPP / 第三军医大学学报

ABSTRACT

Objective To construct the RNAi eukaryotic vector of inhibitory member of the ASPP family(iASPP) gene and observe the interfering effect in Jurkat cell line after the vector transfection.Methods The specific siRNA sequence was designed according to the iASPP sequence in GenBank.The sequence was cloned into PGCsilencer~(TM) H1/Neo/GFP and then sequence analysis was performed.The recombinant plasmid was transfected into Jurkat cell by liposome.The iASPP expression was analyzed by RT-PCR and cell apoptosis was detected by FCM.Results The sequence of templet and specific siRNA was correct by sequence analysis.The iASPP expression in Jurkat cells decreased and the apoptosis rate increased from 11.81% to 33.15% after RNAi transfection.Conclusion The RNAi eukaryotic vector PGCsilencer~(TM)H1/Neo/GFP/RNAi was constructed successfully.The RNAi inhibitory effect on the Jurkat cells is confirmed.The successful construction of iASPP RNAi makes it possible to further study the interaction between iASPP and p53.