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Updated: Apr 26, 2026

Production of Replication-Defective Retrovirus by Transient Transfection of 293T cells
Published on: December 4, 2007
[The effect of retrovirus-mediated hTRT transfection into cultured oral keratinocytes]
Ji-yan Huang1, Wei Liu, Zeng-tong Zhou
1Department of Oral Medicine, Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Stomatology. Shanghai 200011; China.E-mail:coucouh@163.com.
Purpose:
Human telomerase reverse transcriptase (hTRT) was transfected into cultured oral keratinocytes (OKC) mediated by pBABE-tert recombined retrovirus to investigate the effect on OKC lifespan.
Methods:
pBABE-tert recombined retrovirus loaded with hTRT gene was amplified by transfected PT67 cells, and then transfected into cultured OKC in vitro. The positive clones of OKC were separated by puromycin and subcultured. Telomerase activity was analyzed by telomerase PCR-ELISA and PCR-PAGE.
Results:
The hTRT positive clones of OKC showed telomerase expression, with extending lifespan to 8-9 passages.
Conclusions:
The hTRT transfected OKC can prolong doubly lifespan but not be immortalized, which indicates that cellular immortality mechanism is complicated and multi-controled. Telomerase activity is the key for cell immortalization but not the only impact factor.
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