Improved expression by cytomegalovirus promoter/enhancer and behavior of vascular endothelial growth factor gene after myocardial injection of naked DNA.
Direct injection of the vascular endothelial growth factor (VEGF) gene plasmid DNA into the myocardium was shown to induce development of new blood vessels to increase the circulation in the heart of patients with coronary artery diseases. However, such angiogenic gene therapy (via naked DNA) was limited by low level of gene expression. Furthermore, the temporal and spatial characteristics of VEGF gene transfer in the heart are not known. In this study, we demonstrated that a plasmid vector, containing the human cytomegalovirus immediate early (HCMV IE) promoter and enhancer, induces greater expression of gene in the rat heart monitored by gene fused to the chloramphenicol acetyl transferase (CAT) reporter, than four different viral and cellular promoters. Interestingly, expression of VEGF121 protein showed an earlier peak, a shorter duration, and a wider distribution than that of CAT only. Therefore, a plasmid vector with an HCMV IE promoter/enhancer provides clear advantages over other previously developed plasmids. Furthermore, expression profile of VEGF121 gene may provide useful information in the design of angiogenic gene therapy in the heart.
Jeong
Jin Ok
JO
Department of Internal Medicine, Chungnam National University School of Medicine, Daejeon 301-721, Korea.
Byun
Jonghoe
J
Jeon
Eun-Seok
ES
Gwon
Hyeon-Cheol
HC
Lim
Young Shin
YS
Park
Jangwon
J
Yeo
Seon-Ju
SJ
Lee
Young Joo
YJ
Kim
Sunyoung
S
Kim
Duk-Kyung
DK
D000818:Animals; D019677:Artificial Gene Fusion; D015500:Chloramphenicol O-Acetyltransferase / Q000032:analysis / Q000235:genetics; D003587:Cytomegalovirus / Q000235:genetics*; D004279:DNA, Viral / Q000008:administration & dosage* / Q000235:genetics*; D016228:Endothelial Growth Factors / Q000032:analysis / Q000235:genetics*; D004742:Enhancer Elements, Genetic; D015967:Gene Expression Regulation, Viral; D018014:Gene Transfer Techniques; D005814:Genes, Viral; D005822:Genetic Vectors; D036341:Intercellular Signaling Peptides and Proteins / Q000032:analysis / Q000235:genetics*; D008222:Lymphokines / Q000032:analysis / Q000235:genetics*; D008297:Male; D009206:Myocardium / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D051381:Rats; D017207:Rats, Sprague-Dawley; D013997:Time Factors; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D004279:DNA, Viral; D016228:Endothelial Growth Factors; D036341:Intercellular Signaling Peptides and Proteins; D008222:Lymphokines; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors; D015500:Chloramphenicol O-Acetyltransferase