1. [Construction of RNA interfering lentiviral vector of occludin and its role in tight junctions].
- Author
-
Zhu MY, Shen JY, Fei LM, and Chen DY
- Subjects
- Animals, Cell Line, Lentivirus, Mice, Occludin genetics, RNA Interference, Tight Junctions
- Abstract
Objective: To investigate the role of occludin in tight junction (TJ) in vitro., Methods: We constructed RNA interfering lentiviral vectors and transfected them into TM4 cells. Then we detected their inhibitory effect on occuldin by RT-PCR and Western blot and analyzed the role of occuldin in TJ using an in vitro TJ cell model., Results: The pLenti 6.3-EGFP-occludin-miR expression vector was successfully constructed. The results of RT-PCR and Western blot showed that pLenti 6.3-EGFP-occludin-miR-3 significantly inhibited the expression of occludin (P < 0.05), which was remarkably lower than in the blank control and the pLenti 6.3- EGFP transfection group (0.7534 ± 0.089 vs 1.000 and 1.056 ± 0.025, P < 0.05). The expression of occludin was markedly suppressed and the tightness of tight junctions decreased in the TM4 cells transfected with pLenti 6.3-EGFP-occludin-miR-3., Conclusions: The pLenti 6.3-EGFP-occludin-miR expression vector was successfully constructed, and occludin is one of the functional proteins that maintain tight junctions.
- Published
- 2021