DddA 同源物搜索和工程改造扩展了线粒体碱基编辑的序列兼容性。
DddA homolog search and engineering expand sequence compatibility of mitochondrial base editing.
机构信息
Institute of Molecular Medicine, College of Future Technology, Peking University, Beijing, China.
Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China.
出版信息
Nat Commun. 2023 Feb 16;14(1):874. doi: 10.1038/s41467-023-36600-2.
Expanding mitochondrial base editing tools with broad sequence compatibility is of high need for both research and therapeutic purposes. In this study, we identify a DddA homolog from Simiaoa sunii (Ddd_Ss) which can efficiently deaminate cytosine in DC context in double-stranded DNA (dsDNA). We successfully develop Ddd_Ss-derived cytosine base editors (DdCBE_Ss) and introduce mutations at multiple mitochondrial DNA (mtDNA) loci including disease-associated mtDNA mutations in previously inaccessible GC context. Finally, by introducing a single amino acid substitution from Ddd_Ss, we successfully improve the activity and sequence compatibility of DdCBE derived from DddA of Burkholderia cenocepacia (DdCBE_Bc). Our study expands mtDNA editing tool boxes and provides resources for further screening and engineering dsDNA base editors for biological and therapeutic applications.
扩展具有广泛序列兼容性的线粒体碱基编辑工具对于研究和治疗目的都非常有必要。在这项研究中，我们从思茅细辛（Simiaoa sunii）中鉴定出一个 DddA 同源物（Ddd_Ss），它可以在双链 DNA（dsDNA）中有效地将 DC 环境中的胞嘧啶脱氨。我们成功开发了 Ddd_Ss 衍生的胞嘧啶碱基编辑器（DdCBE_Ss），并在多个线粒体 DNA（mtDNA）位点引入突变，包括以前无法触及的 GC 环境中的与疾病相关的 mtDNA 突变。最后，通过从伯克霍尔德菌（Burkholderia cenocepacia）的 DddA 引入一个来自 Ddd_Ss 的单一氨基酸取代，我们成功地提高了 DdCBE 的活性和序列兼容性，该 DdCBE 来自 Ddd_Ss（DdCBE_Bc）。我们的研究扩展了 mtDNA 编辑工具盒，并为进一步筛选和工程 dsDNA 碱基编辑器提供了资源，用于生物和治疗应用。
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