Preventing autosomal-dominant hearing loss in Bth mice with CRISPR/CasRx-based RNA editing.

ABSTRACT: CRISPR/RfxCas13d (CasRx) editing system can specifically and precisely cleave single-strand RNAs, which is a promising treatment for various disorders by downregulation of related gene expression. Here, we tested this RNA-editing approach on Beethoven (Bth) mice, an animal model for human DFNA36 due to a point mutation in Tmc1. We first screened 30 sgRNAs in cell cultures and found that CasRx with sgRNA3 reduced the Tmc1^Bth transcript by 90.8%, and the Tmc1 wild type transcript (Tmc1⁺) by 44.3%. We then injected a newly developed AAV vector (AAV-PHP.eB) based CasRx into the inner ears of neonatal Bth mice, and we found that Tmc1^Bth was reduced by 70.2% in 2 weeks with few off-target effects in the whole transcriptome. Consistently, we found improved hair ce

SUBMITTER: Zheng Z 

PROVIDER: S-EPMC8918553 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature