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
Amino Acids, Peptides, and Proteins | Congenital, Hereditary, and Neonatal Diseases and Abnormalities | Genetics and Genomics | Nervous System Diseases | Neuroscience and Neurobiology | Therapeutics
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.
Huntington’s Disease, Gene Editing, Single Nucleotide Polymorphism, UMCCTS funding
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
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
Human gene therapy
Oikemus S, Pfister EL, 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. (2021). Allele-specific knockdown of mutant HTT protein via editing at coding region SNP heterozygosities. Population and Quantitative Health Sciences Publications. https://doi.org/10.1089/hum.2020.323. Retrieved from https://escholarship.umassmed.edu/qhs_pp/1426
Available for download on Wednesday, August 10, 2022
Amino Acids, Peptides, and Proteins Commons, Congenital, Hereditary, and Neonatal Diseases and Abnormalities Commons, Genetics and Genomics Commons, Nervous System Diseases Commons, Neuroscience and Neurobiology Commons, Therapeutics Commons