1.Correction of the pathogenic mutation in the deafness gene SLC26A4 via prime editor and adenine base editor in vitro
Jiang JIN ; Jineng LYU ; Lei CHEN ; Lili XIE
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2025;60(1):34-41
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.
2.Correction of the pathogenic mutation in the deafness gene SLC26A4 via prime editor and adenine base editor in vitro
Jiang JIN ; Jineng LYU ; Lei CHEN ; Lili XIE
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2025;60(1):34-41
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.
Result Analysis
Print
Save
E-mail