Abstract:

To construct and screen the shRNAsexpressing lentiviral recombinants targeting the rat PTTG gene. MethodsFour groups of shRNA sequences specifically targeting the rPTTG gene were designed and synthesized. These shRNAs were  inserted into lentiviral vectors pGCLGFP. The recombinants′ transfection efficiency was evaluated after infecting NRK cell line with the lentiviral particles. Real time PCR was employed to assess the gene silencing efficacy of these recombinants. ResultsReal time PCR and sequencing showed that shRNAs were correctly inserted into the lentiviral vector and recombinants were constructed successfully. Transfection efficiency of lenviral recombinants was more than 70%. All of these four shRNAs could achieve gene knockdown effect, and 4# shRNA had the most significant gene silence effect among them(＞80%). ConclusionsshRNAsexpressing lentiviral recombinants targeting the rPTTG gene were successfully constructed and screened. Lentivirus can operated as RNAi vectors to achieve effective transfection. 4# vshRNA could inhibit PTTG expression successfully.  

Key words: PTTG； shRNA； Lentivirus