Design of a muscle cell-specific expression vector utilising human vascular smooth muscle alpha-actin regulatory elements.
The facility to direct tissue-specific expression of therapeutic gene constructs is desirable for many gene therapy applications. We describe the creation of a muscle-selective expression vector which supports transcription in vascular smooth muscle, cardiac muscle and skeletal muscle, while it is essentially silent in other cell types such as endothelial cells, hepatocytes and fibroblasts. Specific transcriptional regulatory elements have been identified in the human vascular smooth muscle cell (VSMC) alpha-actin gene, and used to create an expression vector which directs the expression of genes in cis to muscle cells. The vector contains an enhancer element we have identified in the 5' flanking region of the human VSMC alpha-actin gene involved in mediating VSMC expression. Heterologous pairing experiments have shown that the enhancer does not interact with the basal transcription complex recruited at the minimal SV40 early promoter. Such a vector has direct application in the modulation of VSMC proliferation associated with intimal hyperplasia/restenosis.
Keogh
M C
MC
Thrombosis Research Institute, London, UK.
Chen
D
D
Schmitt
J F
JF
Dennehy
U
U
Kakkar
V V
VV
Lemoine
N R
NR
D000199:Actins / Q000235:genetics*; D000818:Animals; D002478:Cells, Cultured; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D008156:Luciferases / Q000235:genetics; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D011817:Rabbits; D051381:Rats; D012045:Regulatory Sequences, Nucleic Acid; D014162:Transfection
D000199:Actins; D008156:Luciferases