Abstract: Objective To analyze the effect of small interfering RNA (siRNA) targeting mouse epididymis-specific colipase-like
(meClps) gene on mouse sperm mobility. Methods The eukaryotic expression vector pDsRed2.0-C1-meClps was constructed
and transfected into NIH-3T3 cells, and the protein expression was detected with anti-meClps serum. Three interfering
sequences targeting meClps (RNAi-251, 224 and 286) were inserted into lentiviral vectors pRNAT-U6.2/lenti, which were
co-transfected with pDsRed2.0-C1-meClps into NIH-3T3 cells. The RNA interfering efficiency was confirmed by
semi-quantitative PCR and Western blotting. The lentivirus, packed with the lentiviral vector with the highest interfering
efficiency, was injected into the caput tissues of mouse epididymis, and its effect on sperm mobility of the cauda epididymis
was evaluated. Results All the 3 lentiviral RNAi vectors targeting meClps could inhibit the mRNA and protein expressions of
meClps, among which pRNAT-U6.2/lenti-RNAi-251 had the highest interfering efficiency. The lentivirus packed with
pRNAT-U6.2/lenti-RNAi-251 significantly reduced the path velocity of cauda sperm after injection into the caput epididymis
of the mice (P<0.05). Conclusion Knock-down meClps expression by lentiviral-mediated RNA interference can lower sperm
mobility of mice.