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17β-Hydroxysteroid dehydrogenase type 7 (Hsd17b7) reverts cholesterol auxotrophy in NS0 cells

Paper ID Volume ID Publish Year Pages File Format Full-Text
25735 43594 2006 12 PDF Available
Title
17β-Hydroxysteroid dehydrogenase type 7 (Hsd17b7) reverts cholesterol auxotrophy in NS0 cells
Abstract

NS0 is a host cell line widely used for the production of recombinant therapeutic proteins. In this work, we investigated the cholesterol-dependent phenotype of NS0 cells. Growth response to different precursors and comparative transcript analyses pointed to deficiency of 17β-hydroxysteroid dehydrogenase type 7 (Hsd17b7) in NS0 cells. Hsd17b7 was previously shown to encode for an enzyme involved in estrogenic steroid biosynthesis. Its recent cloning into a yeast mutant deficient in ERG27 led to its functional characterization as the 3-ketoreductase of the cholesterol biosynthesis pathway. To ascertain that its cholesterol biosynthesis is blocked at the reduction reaction catalyzed by Hsd17b7, we genetically engineered NS0 cells to over express Hsd17b7. The stable transfectants of Hsd17b7 were able to grow independent of cholesterol. The results affirm the role of Hsd17b7 in the cholesterol biosynthesis pathway in mammals. Further, the findings allow for rational engineering of this industrially important cell line to alleviate their cholesterol dependence.

Keywords
Cholesterol biosynthesis; 17β-Hydroxysteroid dehydrogenase type 7; Hsd17b7; NS0; 3-Ketoreductase; Mouse myeloma; Cholesterol auxotrophy; Cell engineering; Mammalian cell culture; Gene expression
First Page Preview
17β-Hydroxysteroid dehydrogenase type 7 (Hsd17b7) reverts cholesterol auxotrophy in NS0 cells
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 121, Issue 2, 24 January 2006, Pages 241–252
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering