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https://hdl.handle.net/2440/59835
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Type: | Journal article |
Title: | Gene silencing of EXTL2 and EXTL3 as a substrate deprivation therapy for heparan sulphate storing mucopolysaccharidoses |
Author: | Kaidonis, X. Liaw, W. Derrick Roberts, A. Ly, M. Anson, D. Byers, S. |
Citation: | European Journal of Human Genetics, 2010; 18(2):194-199 |
Publisher: | Nature Publishing Group |
Issue Date: | 2010 |
ISSN: | 1018-4813 1476-5438 |
Statement of Responsibility: | Xenia Kaidonis, Wan Chin Liaw, Ainslie Derrick Roberts, Marleesa Ly, Donald Anson and Sharon Byers |
Abstract: | Neurological pathology is characteristic of the mucopolysaccharidoses (MPSs) that store heparan sulphate (HS) glycosaminoglycan (gag) and has been proven to be refractory to systemic therapies. Substrate deprivation therapy (SDT) using general inhibitors of gag synthesis improves neurological function in mouse models of MPS, but is not specific to an MPS type. We have investigated RNA interference (RNAi) as a method of targeting SDT to the HS synthesising enzymes, EXTL2 and EXTL3. Multiple shRNA molecules specific to EXTL2 or EXTL3 were designed and validated in a reporter gene assay, with four out of six shRNA constructs reducing expression by over 90%. The three EXTL2-specific shRNA constructs reduced endogenous target gene expression by 68, 32 and 65%, and decreased gag synthesis by 46, 50 and 27%. One EXTL3-specific shRNA construct reduced endogenous target gene expression by 14% and gag synthesis by 39%. Lysosomal gag levels in MPS IIIA and MPS I fibroblasts were also reduced by EXTL2 and EXTL3-specific shRNA. Incorporation of shRNAs into a lentiviral expression system reduced gene expression, and one EXTL2-specific shRNA reduced gag synthesis. These results indicate that deprivation therapy through shRNA-mediated RNAi has potential as a therapy for HS-storing MPSs. |
Keywords: | T-Lymphocytes Lysosomes Humans Mucopolysaccharidoses N-Acetylglucosaminyltransferases Heparitin Sulfate Membrane Proteins Gene Products, gag RNA DNA Primers Transfection Polymerase Chain Reaction Gene Expression Regulation Gene Silencing Genes, Reporter Child |
Rights: | Copyright © 2010, Nature Publishing Group |
DOI: | 10.1038/ejhg.2009.143 |
Published version: | http://dx.doi.org/10.1038/ejhg.2009.143 |
Appears in Collections: | Aurora harvest 5 Molecular and Biomedical Science publications |
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