Center for Genome Engineering, Institute for Basic Science, Daejeon, 34126, Republic of Korea.
Laboratory Animal Resource and Research Center, Korea Research Institute of Bioscience and Biotechnology, Cheongju, 28116, Republic of Korea.
Genome Biol. 2022 Oct 12;23(1):211. doi: 10.1186/s13059-022-02782-z.
We present two methods for enhancing the efficiency of mitochondrial DNA (mtDNA) editing in mice with DddA-derived cytosine base editors (DdCBEs). First, we fused DdCBEs to a nuclear export signal (DdCBE-NES) to avoid off-target C-to-T conversions in the nuclear genome and improve editing efficiency in mtDNA. Second, mtDNA-targeted TALENs (mitoTALENs) are co-injected into mouse embryos to cleave unedited mtDNA. We generated a mouse model with the m.G12918A mutation in the MT-ND5 gene, associated with mitochondrial genetic disorders in humans. The mutant mice show hunched appearances, damaged mitochondria in kidney and brown adipose tissues, and hippocampal atrophy, resulting in premature death.
我们提出了两种方法来提高小鼠中线粒体 DNA（mtDNA）编辑的效率，方法是使用来源于 DddA 的胞嘧啶碱基编辑器（DdCBEs）。首先，我们将 DdCBE 与核输出信号（DdCBE-NES）融合，以避免核基因组中的脱靶 C-to-T 转换，并提高 mtDNA 中的编辑效率。其次，将靶向 mtDNA 的 TALENs（mitoTALENs）共同注射到小鼠胚胎中，以切割未编辑的 mtDNA。我们生成了一个具有 MT-ND5 基因 m.G12918A 突变的小鼠模型，该突变与人类的线粒体遗传疾病有关。突变小鼠表现出弓背外观、肾脏和棕色脂肪组织中线粒体损伤以及海马体萎缩，导致过早死亡。
