Protocol

Cardiac Gene Expression

Volume 366 of the series Methods in Molecular Biology pp 331-342

Adeno-Associated Viral Vector-Delivered Hypoxia-Inducible Gene Expression in Ischemic Hearts

  • Hua SuAffiliated withCardiovascular Research Institute, University of California, San FranciscoDepartment of Medicine, University of California, San Francisco
  • , Yuet Wai KanAffiliated withCardiovascular Research Institute, University of California, San FranciscoDepartment of Medicine, University of California, San FranciscoDepartment of Laboratory Medicine, University of California, San Francisco

* Final gross prices may vary according to local VAT.

Get Access

Abstract

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.

Key Words

Hypoxia-response element hypoxia-inducible gene expression adenoassociated viral vector virus production ischemic heart