关键词: miR-146b, 慢病毒, 人脂肪前体细胞

Abstract: Objective To construct a recombinant lentiviral vector carrying miR-146b and validate its infection efficiency in human pre-adipocytes. Methods The minigene fragment of miR-146b was amplified from human genomic DNA by PCR,then cloned into a lentivirus expression vector.After sequencing validated exactly,the expression plasmid and packaging plasmids Pcgvp,Rev,Vsvg were co-transfected into HEK-293T cells to produce lentivirus; the supernatant containing lentiviral particles was used to infect human preadipocytes,and its infection efficiency and miR-146b overexpression level were respectively tested by fluorescent observation and real-time quantitative PCR. Results MiR-46b lentiviral vector was constructed successfully,and infection efficiency in human pre-adipocytes reached over 85%,the expression level increased nearly four folds. Conclusion We have successfully constructed the lentiviral vector containing miR-146b with high overexpression in infected human pre-adipocytes,which will be useful to the future studying the function of miR-146b in human pre-adipocytes.

Key words: miR-146b, lentivirus, human preadipocytes