CanScript, an 18-Base pair DNA sequence, boosts tumor cell-specific promoter activity: implications for targeted gene therapy.
Gene therapy protocols for the treatment of cancer often employ gene promoter sequences that are known to be over-expressed 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.
Huang
Yu-Hung
YH
Lankenau Institute for Medical Research, Wynnewood, PA, USA.
Cozzitorto
Joseph A
JA
Richards
Nathan G
NG
Eltoukhy
Ahmed A
AA
Yeo
Charles J
CJ
Langer
Robert
R
Anderson
Daniel G
DG
Brody
Jonathan R
JR
Sawicki
Janet A
JA
D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D004247:DNA / Q000235:genetics; D004742:Enhancer Elements, Genetic; D058851:GPI-Linked Proteins / Q000235:genetics*; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D000090204:Mesothelin; D051379:Mice; D058990:Molecular Targeted Therapy / Q000379:methods*; D009374:Neoplasms, Experimental / Q000235:genetics* / Q000628:therapy*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D020080:Tandem Repeat Sequences; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured
D058851:GPI-Linked Proteins; C000715528:Msln protein, mouse; D014157:Transcription Factors; D004247:DNA; D008156:Luciferases; D000090204:Mesothelin
Cancer Res. 2007 Oct 1;67(19):9055-65
17909009
Biotechniques. 1998 Jun;24(6):922-4, 926, 928
9631181
Proc Natl Acad Sci U S A. 2004 Nov 9;101(45):16028-33
15520369
Gene Ther. 2005 Nov;12(21):1573-80
16034457
Adv Exp Med Biol. 2008;622:15-21
18546615
Cell. 1991 May 17;65(4):551-68
1851669
Cancer Biol Ther. 2008 Oct;7(10):1584-90
19039293
Am J Surg Pathol. 2003 Nov;27(11):1418-28
14576474
Proc Natl Acad Sci U S A. 1999 Sep 28;96(20):11531-6
10500211
Mol Ther. 2002 Sep;6(3):415-21
12231179
Hum Pathol. 2003 Jun;34(6):605-9
12827615
Cancer Res. 2009 Aug 1;69(15):6184-91
19643734
Gynecol Oncol. 2005 Nov;99(2):267-77
16061277
Dev Growth Differ. 1995 Aug;37(4):455-459
37281722
Int J Cancer. 1994 Apr 1;57(1):90-7
8150545