Construction of Lentiviral Vector of Syncytin and Establishment of Jurkat Cells with Syncytin Gene Silence.

Objective To construct PGC-LV-syncytin vector and establish Jurkat cell line with syncytin gene knockdown. Methods The targeting syncytin gene sequence was designed by the RNAi software, and were cloned into the PGC-LV vector. The positive clones were confirmed by both PCR and restriction enzyme digestion, finally by DNA sequencing. The lentivirus was then harvested and used to infect Jurkat cells. The infected efficiency was indicated by GFP and detected with fluorescent microscope. The expression level of syncytin gene of Jurkat cell line at mRNA was detected by Real-Time PCR, and then Annexin V-APC-labeled flow cytometry was used to detect cell apoptosis rates. Results A lentiviral vector with syncytin gene interference was constructed, and a Jurkat cell line stably expressing syncytin shRNA for syncytin knockdown was successfully established. Conclusion A lentiviral vector carrying the targeting gene syncytin has been successfully constructed, and a Jurkat cell line stably expressing syncytin shRNA has established with this lentiviral system, the cell line can be as a cell model for further research of role of syncytin in leukemia. [ABSTRACT FROM AUTHOR]