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Updated: Oct 9, 2025

Lentivirus-mediated Genetic Manipulation and Visualization of Olfactory Sensory Neurons in vivo
Published on: May 22, 2011
[Construction of RNA interfering lentiviral vector of occludin and its role in tight junctions]
Mao-Ying Zhu1, Jian-Yun Shen1, Lu-Min Fei1
1School of Food and Biological Engineering, Fuyang Normal University, Fuyang, Anhui 236037, China.
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.
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