A comparison of adenovirally delivered molecular methods to inhibit Na+/Ca2+ exchange.
The ability to use molecular biology tools to down-regulate Na+/Ca2+ exchanger (NCX) expression will allow us to better understand the regulation of Ca(i)2+ and contractility in heart. Three different techniques to deplete NCX expression were compared: short hairpin RNA (shRNA), antisense RNA and exchanger inhibitory peptide expression via adenoviral transfection. Our results demonstrate that the most efficient method to deplete NCX expression and activity from cardiomyocytes is shRNA. It is also possible to replace the endogenous NCX with alternative isoforms or mutant forms of the NCX. Adenovirally delivered shRNA is an efficient tool for the study of the NCX and could be adapted for many other cardiac proteins.
Hurtado
Cecilia
C
Institute of Cardiovascular Sciences, St. Boniface General Hospital Research Centre, and Departments of Physiology, Faculties of Medicine and Pharmacy, University of Manitoba, Winnipeg, MB Canada R2H 2A6.
Wigle
Jeffrey T
JT
Dibrov
Elena
E
Maddaford
Thane G
TG
Pierce
Grant N
GN
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D016373:DNA, Antisense / Q000235:genetics; D004285:Dogs; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000494:pharmacology*; D051381:Rats; D019831:Sodium-Calcium Exchanger / Q000037:antagonists & inhibitors* / Q000187:drug effects / Q000235:genetics / Q000378:metabolism
D016373:DNA, Antisense; D034741:RNA, Small Interfering; D019831:Sodium-Calcium Exchanger; C110374:sodium-calcium exchanger 1