Construction of targeted lentiviral vector shRNA of CD47 gene and its effect of CD47 gene silence in Hep-2 cells

【Objective】To construct short hairpin RNA interference lentiviral vector of CD47 gene and to evaluate their inhibitory effect in Hep-2 cells. 【Methods】The CD47 target specific oligonucleotide sequence was designed, synthesized and connected into the pGCSIL-GFP vector digested, 293T cells were packaged to produce the lentivirus, and then transfected with Hep-2 cells. The inhibitory effect on CD47 gene was tested by the real-time quantitative PCR and Western Blotting. 【Results】The PCR and DNA sequencing of positive clones demonstrated that the lentivirus shRNA vector of CD47 gene was constructed successfully. The real-time quantitative PCR and Western Blotting confirmed that the vector could suppress the expression level of CD47 at mRNA and protein levels. 【Conclusions】CD47-shRNA lentivirus vector has been constructed successfully, and our recombined lentivirus vector will be a good model for further study and utilization on gene function research about CD47.