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 Blood, 1 December 2006, Vol. 108, No. 12, pp. 3757-3760.
Prepublished online as a Blood First Edition Paper on August 10, 2006; DOI 10.1182/blood-2006-02-004671.

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HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
Brief report

siRNA silencing of calumenin enhances functional factor IX production

Nadeem Wajih, Susan M. Hutson, and Reidar Wallin

From the Departments of Internal Medicine and Biochemistry, Wake Forest University School of Medicine, Winston-Salem, NC.

To improve production of functional fully {gamma}-carboxylated recombinant human clotting factor IX (r-hFIX), cell lines stably overexpressing r-hFIX have been engineered to also overexpress proteins of the {gamma}-carboxylation system. Here we demonstrate that siRNA silencing of calumenin, an inhibitor of the {gamma}-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.


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