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An shRNA silencing a non‐toxic transgene reduces nutrient consumption and increases production of adenoviral vectors in a novel packaging cell
Author(s) -
Wang Lifu,
Qi Xiaoguang,
Shen Ruizhe,
Sun Ying,
Tuveson David A.
Publication year - 2009
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.21679
Subject(s) - small hairpin rna , transgene , gene silencing , viral vector , biology , rna interference , genetic enhancement , cell culture , microbiology and biotechnology , gene , rna , biochemistry , recombinant dna , gene knockdown , genetics
During production of an adenoviral vector in a packaging cell, transgene is expressed concomitantly with the adenoviral gene. Depending on the protein encoded by the transgene, the yield of an adenoviral vector can be reduced or blocked as a result of transgene expression in the packaging cell. We tested the effect of a short hairpin RNA (shRNA) inhibiting the expression of a transgene encoding hIcon, a therapeutic molecule that selectively destroys pathological angiogenic blood vessels, on the yield of an adenoviral vector containing the transgene. The results showed that the yield of infectious adenoviral vector particles was increased about 10‐fold in a novel packaging cell with stable production of an shRNA that can silence the transgene, as compared to the yield in standard packaging line, and the consumption of nutrient in the novel packaging cell line is decreased due to silence of adenoviral transgene expression. The study indicates that shRNA can increase the production of adenoviral vectors by silencing transgene and reducing nutrient consumption of the novel packaging cell. The use of shRNA silencing expression of transgenes encoding therapeutic molecules can reduce the time and cost of producing adenoviral vectors for clinical use. J. Cell. Physiol. 219: 365–371, 2009. © 2008 Wiley‐Liss, Inc.