TY - JOUR
T1 - CanScript, an 18-Base pair DNA sequence, boosts tumor cell-specific promoter activity
T2 - Implications for targeted gene therapy
AU - Huang, Yu Hung
AU - Cozzitorto, Joseph A.
AU - Richards, Nathan G.
AU - Eltoukhy, Ahmed A.
AU - Yeo, Charles J.
AU - Langer, Robert
AU - Anderson, Daniel G.
AU - Brody, Jonathan R.
AU - Sawicki, Janet A.
N1 - Funding Information:
We wish to thank Jiping Chen for technical assistance. We also thank Scott E. Kern (Johns Hopkins University, Baltimore, MD) for kindly donating, and allowing the use of, the original CanScript clone, and for helpful suggestions and discussion. This work was supported by the National Institutes of Health (CA132091 to J.A.S., EB000244 to R.L.); the Teal Ribbon Ovarian Cancer Research Foundation, Inc. to J.A.S.; Wawa, Inc. Corporate Charities to J.A.S.; and the Red Bra Ladies to J.B. and J.A.S.; Supported by Grant #IRG-08-060-01 from the American Cancer Society (J.B.) and a PanCAN Career development Award, American Association of Cancer Research (J.B.).
PY - 2010/11/1
Y1 - 2010/11/1
N2 - Gene therapy protocols for the treatment of cancer often employ gene promoter sequences that are known to be overexpressed in specific tumor cell types relative to normal cells. These promoters, while specific, are often weakly active. It would be desirable to increase the activity of such promoters, while at the same time retain specificity, so that the therapeutic gene is more robustly expressed. Using a luciferase reporter DNA construct in both in vitro cell transfection assays and in vivo mouse tumor models, we have determined that in the absence of any other DNA sequence, a previously identified 18-base pair enhancer sequence called CanScript, lying upstream of the MSLN gene, has ∼25% of the promoter activity of CAG, a very strong non-specific promoter/enhancer, in tumor cells in which MSLN is highly expressed. Furthermore, tandem repeat copies of CanScript enhance transcription in a dose-dependent manner and, when coupled with promoter sequences that are active in tumor cells, increase promoter activity. These findings suggest that the incorporation of CanScript into gene constructs may have application in enhancing activity of promoters used in cancer-targeting gene therapy strategies, thereby improving therapeutic efficacy.
AB - Gene therapy protocols for the treatment of cancer often employ gene promoter sequences that are known to be overexpressed in specific tumor cell types relative to normal cells. These promoters, while specific, are often weakly active. It would be desirable to increase the activity of such promoters, while at the same time retain specificity, so that the therapeutic gene is more robustly expressed. Using a luciferase reporter DNA construct in both in vitro cell transfection assays and in vivo mouse tumor models, we have determined that in the absence of any other DNA sequence, a previously identified 18-base pair enhancer sequence called CanScript, lying upstream of the MSLN gene, has ∼25% of the promoter activity of CAG, a very strong non-specific promoter/enhancer, in tumor cells in which MSLN is highly expressed. Furthermore, tandem repeat copies of CanScript enhance transcription in a dose-dependent manner and, when coupled with promoter sequences that are active in tumor cells, increase promoter activity. These findings suggest that the incorporation of CanScript into gene constructs may have application in enhancing activity of promoters used in cancer-targeting gene therapy strategies, thereby improving therapeutic efficacy.
KW - Canscript
KW - Gene promoter
KW - Mesothelin
KW - Targeted gene therapy
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U2 - 10.4161/cbt.10.9.13234
DO - 10.4161/cbt.10.9.13234
M3 - Article
C2 - 20798601
AN - SCOPUS:78549250783
SN - 1538-4047
VL - 10
SP - 878
EP - 884
JO - Cancer Biology and Therapy
JF - Cancer Biology and Therapy
IS - 9
ER -