Correction of the pathogenic mutation in the deafness gene SLC26A4 via prime editor and adenine base editor in vitro
10.3760/cma.j.cn115330-20240420-00221
- VernacularTitle:先导编辑和腺嘌呤碱基编辑体外纠正耳聋基因 SLC26A4致病变异
- Author:
Jiang JIN
1
;
Jineng LYU
;
Lei CHEN
;
Lili XIE
Author Information
1. 温州市人民医院 温州医科大学温州市第三临床学院 温州市妇幼保健院检验科，温州 325027
- Publication Type:Journal Article
- Keywords:
Hearing loss, sensorineural;
Mutation;
SLC26A4;
Prime editor;
Adenine base editor;
In vitro
- From:
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
2025;60(1):34-41
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the feasibility of in vitro prime editor (PE) and adenine base editor (ABE) for correction the pathogenic variant of the human deafness gene SLC26A4 c.1229C>T. Methods:From March 2023 to April 2024, prime editing guide RNA (pegRNA) expression vectors as well as single guide RNA (sgRNA) were designed and constructed for the SLC26A4 c.1229C>T variant, and the feasibility of correction was performed in the HEK293T mutation model, the correction efficiency was analyzed by deep sequencing. Results:A mutant cell model of SLC26A4 c.1229C>T was successfully established. Correction was achieved in the SLC26A4 c.1229C>T mutant cell model using PE and ABE8e. Deep sequencing analysis revealed the correction efficiencies of (31.89±0.77)% and (41.07±2.28)%, respectively. Conclusion:In this study, a new base correction strategy based on the human deafness gene SLC26A4 is proposed, which provides a viable reference for gene therapy of deafness caused by SLC26A4 gene mutation.
