Adeno-associated viral vector-delivered hypoxia-inducible gene expression in ischemic hearts.
This chapter describes a system using adeno-associated viral (AAV) vector to deliver hypoxia-inducible gene expression to ischemic hearts. The hypoxia induction of gene expression in this system is based on the accumulation of hypoxia-inducible factor-1 (HIF-1) in ischemic hearts and the use of hypoxia-response element (HRE) identified from the enhancers of genes, the expression of which can be induced by hypoxia. The methods of plasmid and AAV vector construction for hypoxia-inducible gene expression, viral vector production and purification, and viral titer determination are described. This chapter also illustrates the methods that can be used to test hypoxia-inducible gene expression in vitro and in vivo, including hypoxia treatment of cultured cells, generation of murine ischemic heart models, and analysis of gene expression.
Su
Hua
H
Cardiovascular Research Institute/Department of Medicine, University of California, San Francisco, USA.
Kan
Yuet Wai
YW
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics*; D004921:Erythropoietin / Q000235:genetics; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics* / Q000378:metabolism; D007763:Lac Operon / Q000502:physiology; D051379:Mice; D008969:Molecular Sequence Data; D017202:Myocardial Ischemia / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D020218:Response Elements; D014157:Transcription Factors; D014162:Transfection; D042442:Vascular Endothelial Growth Factors / Q000235:genetics / Q000378:metabolism
C497442:HIF1A protein, human; C497443:Hif1a protein, mouse; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D014157:Transcription Factors; D042442:Vascular Endothelial Growth Factors; D004921:Erythropoietin