Lentiviral vector-mediated RNA interference of mouse epididymis-specific meClps gene lowers mouse sperm mobility.
- Author:
Zizhen LIAN
1
;
Zuowu CAO
;
Ran CHEN
;
Lei CHEN
;
Yingzi XUE
;
Junwen QIN
;
Xufeng QI
;
Chunxue ZHANG
;
Yanhong YU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Epididymis; Gene Targeting; Genetic Vectors; Lentivirus; Male; Mice; NIH 3T3 Cells; RNA Interference; RNA, Messenger; RNA, Small Interfering; Sperm Motility; Spermatozoa; Transfection
- From: Journal of Southern Medical University 2014;34(9):1359-1364
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo analyze the effect of small interfering RNA (siRNA) targeting mouse epididymis-specific colipase-like (meClps) gene on mouse sperm mobility.
METHODSThe 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.
RESULTSAll 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).
CONCLUSIONKnock-down meClps expression by lentiviral-mediated RNA interference can lower sperm mobility of mice.
