Abstract： Objective To construct the recombinant RNA interference vector targeting Atp6i and T cell immune response cDNA7 (TIRC7),and to prepare its recombinant AAV particle in order to provide evidence for gene therapy for diseases using the recombinant RNA interference vector in vivo and in vitro.Methods Sequences of small interference iRNA targeting the common region of Atp6i and TIRC7 genes were designed and connected to the pAAV.H1 vector linearized by enzyme digestion of Xba Ⅰ and Bgl Ⅱ and then transformed into DH5α where plasmid was identified using enzyme digestion for sequencing analysis.The identified siRNA sequences,pAAV-RC and pHelper were used to transfect AAV-293 cells by calcium phosphate precipitation method for recombinant AAV.H1Atp6i particle,with phosphate buffer solution(PBS)used in blank control group and AAV.H1Luc in negative control group.The transfection efficiency was then observed by fluorescence microscopy.Western blotting was used to detect the effects of the recombinant AAV.H1Atp6i particle on TIRC7 protein.Results The recombinant siRNA targeting Atp6i and TIRC7 was successfully constructed.Furthermore,AAV-293 cells were successfully transfected by the recombinant particle,pAAV-RC and pHelper,with the transfection efficiency being almost 100％.And the recombinant AAV.H1Atp6i particle was successfully packed and obtained.Western blotting showed the expressions of TIRC7 protein in blank control,negative control and AAV.H1Atp6i treatment groups,with the molecular weight being 75 000.Nevertheless,the TIRC7 expression was decreased by 80％ in AAV.H1Atp6i treatment group as compared with that in blank control and negative control groups (0.271±0.072 vs 0.988±0.042 and 0.992±0.053,all P＜0.05).Conclusion The recombinant siRNA targeting Atp6i and TIRC7 and recombinant AAV.H1Atp6i particle are both successfully obtained,which may exhibit interference on TIRC7 expression.