Targeted A-to-G base editing in human mitochondrial DNA with programmable deaminases
Targeted A-to-G base editing in human mitochondrial DNA with programmable deaminases
复制标题
DOI：
10.1016/j.cell.2022.03.039
复制
发表时间：
2022-05-12
期刊：
影响因子：
64.5
通讯作者：
Kim, Jin-Soo
中科院分区：
文献类型：
--
作者：
Cho, Sung-Ik;Lee, Seonghyun;Kim, Jin-Soo
线粒体DNA（mtDNA）编辑为细胞系和动物中线粒体遗传疾病的疾病建模以及未来治疗这些疾病铺平了道路。然而，使得能够进行mtDNA编辑的细菌胞苷脱氨酶DddA衍生的胞嘧啶碱基编辑器（DdCBE）在很大程度上限于5 '-TC背景下的C至T转化（例如，TC至TT转化），适合于仅产生所有可能的转换（嘌呤至嘌呤和嘧啶至嘧啶）突变的1/8。在这里，我们提出了转录激活因子样效应子（TALE）连接的脱氨酶（TALEDS），由定制设计的TALE DNA结合阵列、源自新洋葱伯克霍尔德菌的催化受损的全长DddA变体或分裂DddA组成，以及来自大肠杆菌的工程化脱氧腺苷脱氨酶。coil TadA蛋白，其在人类线粒体中诱导靶向A-to-G编辑。定制设计的TALED在人类细胞中非常高效，在各种线粒体基因中总共17个靶位点催化A到G的转换，编辑频率高达49%。
Mitochondrial DNA (mtDNA) editing paves the way for disease modeling of mitochondrial genetic disorders in cell lines and animals and also for the treatment of these diseases in the future. Bacterial cytidine deaminase DddA-derived cytosine base editors (DdCBEs) enabling mtDNA editing, however, are largely limited to C-to-T conversions in the 5'-TC context (e.g., TC-to-TT conversions), suitable for generating merely 1/8 of all possible transition (purine-to-purine and pyrimidine-to-pyrimidine) mutations. Here, we present transcription-activator-like effector (TALE)-linked deaminases (TALEDs), composed of custom-designed TALE DNA-binding arrays, a catalytically impaired, full-length DddA variant or split DddA originated from Burkholderia cenocepacia, and an engineered deoxyadenosine deaminase derived from the E. coil TadA protein, which induce targeted A-to-G editing in human mitochondria. Custom-designed TALEDs were highly efficient in human cells, catalyzing A-to-G conversions at a total of 17 target sites in various mitochondria' genes with editing frequencies of up to 49%.