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Open Access Green as soon as Postprint is submitted to ZB.
Functional analysis of cholesterol biosynthesis by RNA interference.
J. Steroid Biochem. Mol. Biol. 104, 105-109 (2007)
Inborn errors of cholesterol biosynthesis caused by dysfunctionality of single enzymes are known to cause severe malformation syndromes like X-linked chondrodysplasia punctata (CDPX2), CHILD syndrome or Smith-Lemli-Opitz-syndrome (SLOS). In this study we established the method of RNA interference (RNAi) for analyzing the molecular mechanisms underlying disrupted cholesterol biosynthesis. For different genes involved in the cholesterol biosynthesis pathway-NAD(P) dependent steroid dehydrogenase-like (NSDHL), 17-beta hydroxysteroid dehydrogenase type 7 (HSD17B7) and emopamil binding protein (EBP)-shRNA sequences were designed and tested for their effectiveness. For a better comparability of the experiments and to avoid different transfection efficiencies, examined shRNA sequences which reached a knock down of at least 80% were stably transfected in a HeLa cell line with a tetracycline-regulated expression (HeLa T-REx). These stable transfected cell lines represent novel tools for the analysis of cholesterol biosynthesis.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Cholesterol biosynthesis; RNA interference; HSD17B7; NSDHL; EBP
ISSN (print) / ISBN
0960-0760
e-ISSN
0960-0760
Quellenangaben
Volume: 104,
Issue: 3-5,
Pages: 105-109
Publisher
Elsevier
Non-patent literature
Publications
Reviewing status
Peer reviewed
Institute(s)
Molekulare Endokrinologie und Metabolismus (MEM)