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Comparison of the specificity of three endothelial‐specific promoters: Flt1, ICAM2 and vWF
Author(s) -
Lei Jing,
Kanno Shinichi,
Wang Zifa,
Billiar Timothy R
Publication year - 2006
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/fasebj.20.5.a1104-d
Subject(s) - promoter , microbiology and biotechnology , transduction (biophysics) , staining , adventitia , biology , ex vivo , lac operon , gene expression , gene , in vivo , biochemistry , anatomy , genetics
Endothelial‐specific targeting is an attractive topic in vascular gene therapy. To find a suitable promoter for endothelial‐specific gene expression, the specificity of three endothelial promoters was compared. These included fms ‐like tyrosine kinase‐1 (Flt‐1, −748~284bp), intercellular adhesion molecule‐2 (ICAM‐2, −294~44bp), and von Willebrand factor (vWF, −487~247bp) promoter. The three promoters were constructed in an adenovirus as AdFlt1‐lacZ, AdICAM2‐lacZ and AdvWF‐lacZ, respectively. Methods In an ex vivo transduction model, rat aortas, cardiac muscle and skeletal muscle were transduced by the three adenoviral vectors for 2hrs, then cultured in fresh media for 72 hours. After that X‐gal staining was performed to detect the lacZ gene expression. Results In the AdFlt1‐lacZ transduced aorta, about 10% of the endothelial cells and 8% of the adventitia cells showed X‐gal staining, but no staining was evident in the smooth muscle cells. A few of the cardiac muscle and skeletal muscle cells also showed X‐gal staining. In the AdICAM2‐lacZ transduced tissues, 7% of the endothelial cells and 2% of the adventitia cells showed X‐gal staining, while few other types of the cells exhibited positive staining. In the AdvWF‐lacZ transduced tissues, few cells showed X‐gal staining. Conclusion Both the ICAM2 and Flt1 promoters exhibited good endothelial activity, and the ICAM2 promoter exhibited better specificity than the Flt1 promoter. Both show promise for endothelial‐specific targeting in gene therapy. The specificity of these promoters will be further investigated in an in vivo transduction study.