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An enhancer region of the ALK1 gene resides in its intron 2
Author(s) -
Suzuki Rumi,
Yonenaga Yoshikuni,
Lan Fei,
Seki Tsugio
Publication year - 2009
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.23.1_supplement.634.10
Subject(s) - enhancer , biology , reporter gene , gene , intron , regulatory sequence , conserved sequence , genetics , gene expression , microbiology and biotechnology , peptide sequence
Expression of activin receptor‐like kinase 1 (ALK1) gene is undetectable in most of the organs in adults; however, it is specifically induced in arteries feeding ischemic lesions. This gene, with its unique expression pattern, might be exploited to aid in the delivery of a therapeutic peptide to downstream ischemic tissues. Previously, we generated a shortened 4.8 kb Alk1PIB (Promoter‐Intron 2B) regulatory fragment that recapitulates the endogenous mouse ALK1 regulatory activity. In order to elucidate the regulatory mechanisms further, we utilized evolutionarily conserved region (ECR) analysis to identify highly conserved regulatory regions in this study. An alignment of ALK1 gene sequences revealed a 152 bp sequence conserved among 11 mammals within the intron 2B region. To test the regulatory function of this 152‐bp coreECR sequence, recombinant adenoviruses expressing fluorescent reporter were created. Compared to an Alk1PIB virus, the coreECR‐deleted Alk1PIB(d1) virus showed significantly reduced reporter gene expression in cultured arterial endothelial cells, indicating an enhancer function of this sequence. This approach should help us further engineer the ALK1 regulatory region to create short and strong fragment that may be utilized in future gene therapy of various disorders including malignant tumors and ischemic heart diseases. This work was supported by the American Heart Association.