Allele-Specific Knockdown of Mutant Huntingtin Protein via Editing at Coding Region Single Nucleotide Polymorphism Heterozygosities
| Autor: | Ellen Sapp, Rachael Miller, Kathryn Chase, Sarah R. Oikemus, Eric Mick, Lihua Julie Zhu, Michael H. Brodsky, Miguel Sena-Esteves, Edward Hudgens, Akanksh Chaudhary, Lori A. Kennington, Scot A. Wolfe, Neil Aronin, Marian DiFiglia, Edith L. Pfister |
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| Rok vydání: | 2022 |
| Předmět: |
Genetics
Huntingtin Protein Transgene Mutant Single-nucleotide polymorphism Biology medicine.disease Polymorphism Single Nucleotide Mice Huntington Disease Huntington's disease medicine Animals Molecular Medicine Coding region Allele Trinucleotide repeat expansion Indel Molecular Biology Research Articles Alleles |
| Zdroj: | Hum Gene Ther |
| ISSN: | 1557-7422 1043-0342 |
| DOI: | 10.1089/hum.2020.323 |
| Popis: | Huntington's disease (HD) is a devastating, autosomal dominant neurodegenerative disease caused by a trinucleotide repeat expansion in the huntingtin (HTT) gene. Inactivation of the mutant allele by clustered regularly interspaced short palindromic repeats (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 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. |
| Databáze: | OpenAIRE |
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