Streamlined process for effective and strand-selective mitochondrial base editing using mitoBEs

Abstract

Mitochondrial base editing tools hold great promise for the investigation and treatment of mitochondrial diseases. Mitochondrial DNA base editors (mitoBEs) integrate a programmable transcription-activator-like effector (TALE) protein with single-stranded DNA deaminase (TadA8e-V106W, APOBEC1, etc.) and nickase (MutH, Nt.BspD6I(C), etc.) to achieve heightened precision and efficiency in mitochondrial base editing. This innovative mitochondrial base editing tool exhibits a number of advantages, including strand-selectivity for editing, high efficiency, and the capacity to perform diverse types of base editing on the mitochondrial genome by employing various deaminases. In this context, we provide a detailed experimental protocol for mitoBEs to assist others in achieving proficient mitochondrial base editing.

Graphical abstract

Keywords

Mitochondrial DNA base editors (mitoBEs) / Transcription-activator-like effector (TALE) / Deaminase / Nickase

Acknowledgements

This project was supported by funds from the Beijing Municipal Science & Technology Commission (Z181100001318009), the National Science Foundation of China (NSFC31930016), the Beijing Advanced Innovation Center for Genomics at Peking University and the Peking-Tsinghua Center for Life Sciences (to W. Wei), and the Fellowship of China Postdoctoral Science Foundation (to Z. Yi).

RIGHTS & PERMISSIONS

2024 The Author(s)