Submitted February 21, 2006
Accepted July 3, 2006
siRNA Sinencing of Calumenin Enhances Functional Factor IX Production
Nadeem Wajih, Susan M Hutson, and Reidar Wallin*
Department of Internal Medicine, Wake Forest University School of Medicine
Department Biochemistry, Wake Forest University School of Medicine
Department of Internal Medicine, Wake Forest Univ. Sch. Med.
* Corresponding author; email: rwallin{at}wfubmc.edu.
Abstract
To improve production of functional fully ã -carboxylated recombinant human clotting factor IX (r-hFIX), cell lines stably overexpressing r-hFIX have been engineered to also overexpress proteins of the ã -carboxylation system. Here we demonstrate that siRNA silencing of calumenin, an inhibitor of the ã -carboxylation system, enhances production of functional r-hFIX produced by engineered BHK21 cells. The production yield of functional r-hFIX was 80% in engineered cells where calumenin had been silenced 78%. We propose that this high yield expression system can easily be adapted to overproduce functional forms of all members of the vitamin K-dependent protein family.
