UMMS Affiliation

Department of Molecular, Cell and Cancer Biology; Department of Medicine; Department of Population and Quantitative Health Sciences; Horae Gene Therapy Center; Graduate School of Biomedical Sciences

Publication Date

2021-08-10

Document Type

Article Postprint

Disciplines

Amino Acids, Peptides, and Proteins | Congenital, Hereditary, and Neonatal Diseases and Abnormalities | Genetics and Genomics | Nervous System Diseases | Neuroscience and Neurobiology | Therapeutics

Abstract

Huntington's disease (HD) is a devasting, autosomal dominant neurodegenerative disease caused by a trinucleotide repeat expansion in the HTT gene. Inactivation of the mutant allele by CRISPR-Cas9 based gene editing offers a possible therapeutic approach for this disease, but permanent disruption of normal HTT function might compromise adult neuronal function. Here, we use a novel HD mouse model to examine allele-specific editing of mutant HTT (mHTT), with a BAC97 transgene expressing mHTT and a YAC18 transgene expressing normal HTT. We achieve allele-specific inactivation of HTT by targeting a protein coding sequence containing a common, heterozygous single nucleotide polymorphism (SNP). The outcome is a marked and allele-selective reduction of mutant HTT (mHTT) protein in a mouse model of HD. Expression of a single CRISPR-Cas9 nuclease in neurons generated a high frequency of mutations in the targeted HD allele that included both small insertion/deletion (InDel) mutations and viral vector insertions. Thus, allele-specific targeting of InDel and insertion mutations to heterozygous coding region SNPs provides a feasible approach to inactivate autosomal dominant mutations that cause genetic disease.

Keywords

Huntington’s Disease, Gene Editing, Single Nucleotide Polymorphism

Rights and Permissions

© Mary Ann Liebert, Inc. 2021. PDF of authors' peer-reviewed accepted manuscript posted with a 12-month embargo as allowed by the publisher's self-archiving policy at https://home.liebertpub.com/authors/policies/152#self-archiving. Final publication is available from Mary Ann Liebert, Inc., publishers https://doi.org/10.1089/hum.2020.323.

DOI of Published Version

10.1089/hum.2020.323

Source

Oikemus SR, Pfister E, Sapp E, Chase KO, Kennington LA, Hudgens E, Miller R, Zhu LJ, Chaudhary A, Mick EO, Sena-Esteves M, Wolfe SA, DiFiglia M, Aronin N, Brodsky MH. Allele-specific knockdown of mutant HTT protein via editing at coding region SNP heterozygosities. Hum Gene Ther. 2021 Aug 10. doi: 10.1089/hum.2020.323. Epub ahead of print. PMID: 34376056. Link to article on publisher's site

Journal/Book/Conference Title

Human gene therapy

PubMed ID

34376056

Related Resources

Link to Article in PubMed

Available for download on Wednesday, August 10, 2022

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