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Analysis of promoters and expression‐targeted gene therapy optimization based on doubling time and transfectability
Author(s) -
Dobek Georgina L.,
Zhang Xiujuan,
Balazs Daniel A.,
Godbey W. T.
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.11-185421
Subject(s) - survivin , promoter , gene expression , biology , enhancer , gene , reporter gene , genetic enhancement , endogeny , cancer research , microbiology and biotechnology , genetics , endocrinology
ABSTRACT Genes under the control of the cyclooxygenase‐2 (Cox‐2), human epidermal growth factor receptor 2 (Her‐2), and survivin promoters were constructed and delivered to murine and human carcinoma cells. It was found that P Cox‐2‐driven reporter expression was strong and correlated well with endogenous Cox‐2 levels, while P Her‐2 and P survivin yielded poor results, consistent with the three distinct expression mechanisms used by cancer cells to overexpress the endogenous versions of the selected genes. The P Cox‐2 was then used to drive the expression of caspase genes both in vitro and in vivo to bring about targeted apoptosis of carcinoma cells successfully. The results led to the following conclusions. 1 ) When selecting a promoter/enhancer for expression‐targeted gene delivery, it is not enough to perform a microarray on some tumor tissue and select the control element associated with the greatest amount of gene up‐regulation vs. normal controls. The mechanism of expression for the particular gene should be taken into account to prevent lengthy and costly research trials. 2 ) When overexpression is due to activator binding, a predictive model based on endogenous gene expression levels, overall cell transfectability, and cell doubling rates can be used to predict expression‐targeted gene delivery outcomes with significant accuracy.—Dobek, G. L., Zhang, X., Balazs, D. A., Godbey, W. T. Analysis of promoters and expression‐targeted gene therapy optimization based on doubling time and transfectability. FASEB J. 25, 3219‐3228 (2011). www.fasebj.org