利用 mtZFNs 对小鼠线粒体 DNA 异质性的操作。

Manipulation of Murine Mitochondrial DNA Heteroplasmy with mtZFNs.

机构信息

Medical Research Council Mitochondrial Biology Unit, University of Cambridge, Cambridge, UK.

出版信息

Methods Mol Biol. 2023;2615:329-344. doi: 10.1007/978-1-0716-2922-2_23.

Abstract

Mouse models of mitochondrial DNA mutations hold promise in the development and optimization of mitochondrial gene therapy technology and for gathering pre-clinical data prior to human trials. Their suitability for this purpose stems from the high similarity of human and murine mitochondrial genomes and the increasing availability of rationally designed AAV vectors capable of selectively transducing murine tissues. Our laboratory routinely optimizes mitochondrially targeted zinc finger nucleases (mtZFNs), the compactness of which makes them highly suitable for downstream AAV-based in vivo mitochondrial gene therapy. This chapter discusses the necessary precautions for the robust and precise genotyping of the murine mitochondrial genome as well as the optimization of mtZFNs intended for subsequent use in vivo.

摘要

线粒体 DNA 突变的小鼠模型在开发和优化线粒体基因治疗技术以及在进行人体试验之前收集临床前数据方面具有广阔的应用前景。它们适用于此目的，是因为人类和鼠类线粒体基因组的高度相似性，以及越来越多的能够选择性转导鼠类组织的理性设计的 AAV 载体的可用性。我们实验室经常优化靶向线粒体的锌指核酸酶（mtZFNs），其紧凑性使其非常适合用于基于 AAV 的体内线粒体基因治疗的下游应用。本章讨论了稳健和精确的鼠类线粒体基因组基因分型以及优化用于体内后续使用的 mtZFNs 所需的必要预防措施。