Safe and Efficient Silencing with a Pol II, but Not a Pol lII, Promoter Expressing an Artificial miRNA Targeting Human Huntingtin
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Authors
Pfister, Edith L.Chase, Kathryn O.
Sun, Huaming
Kennington, Lori A.
Conroy, Faith
Johnson, Emily S.
Miller, Rachael
Borel, Florie
Aronin, Neil
Mueller, Christian
UMass Chan Affiliations
Department of Pediatrics, Division of PulmonologyRNA Therapeutics Institute
Horae Gene Therapy Center
Department of Medicine
Document Type
Journal ArticlePublication Date
2017-06-16Keywords
AAVHTT
Huntington’s disease
RNAi
gene therapy
huntingtin
miRNA
shRNA
Genetics and Genomics
Nervous System Diseases
Therapeutics
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Show full item recordAbstract
Huntington's disease is a devastating, incurable neurodegenerative disease affecting up to 12 per 100,000 patients worldwide. The disease is caused by a mutation in the Huntingtin (Htt) gene. There is interest in reducing mutant Huntingtin by targeting it at the mRNA level, but the maximum tolerable dose and long-term effects of such a treatment are unknown. Using a self-complementary AAV9 vector, we delivered a mir-155-based artificial miRNA under the control of the chicken beta-actin or human U6 promoter. In mouse brain, the artificial miRNA reduced the human huntingtin mRNA by 50%. The U6, but not the CbetaA promoter, produced the artificial miRNA at supraphysiologic levels. Embedding the antisense strand in a U6-mir-30 scaffold reduced expression of the antisense strand but increased the sense strand. In mice treated with scAAV9-U6-mir-155-HTT or scAAV9-CbetaA-mir-155-HTT, activated microglia were present around the injection site 1 month post-injection. Six months post-injection, mice treated with scAAV9-CbetaA-mir-155-HTT were indistinguishable from controls. Those that received scAAV9-U6-mir-155-HTT showed behavioral abnormalities and striatal damage. In conclusion, miRNA backbone and promoter can be used together to modulate expression levels and strand selection of artificial miRNAs, and in brain, the CbetaA promoter can provide an effective and safe dose of a human huntingtin miRNA.Source
Mol Ther Nucleic Acids. 2017 Jun 16;7:324-334. doi: 10.1016/j.omtn.2017.04.011. Epub 2017 Apr 14. Link to article on publisher's siteDOI
10.1016/j.omtn.2017.04.011Permanent Link to this Item
http://hdl.handle.net/20.500.14038/43582PubMed ID
28624208Related Resources
Link to Article in PubMedDistribution License
http://creativecommons.org/licenses/by-nc-nd/4.0/ae974a485f413a2113503eed53cd6c53
10.1016/j.omtn.2017.04.011
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Except where otherwise noted, this item's license is described as http://creativecommons.org/licenses/by-nc-nd/4.0/