The FusX TALE Base Editor (FusXTBE) for Rapid Mitochondrial DNA Programming of Human Cells In Vitro and Zebrafish Disease Models In Vivo.
The FusX TALE Base Editor (FusXTBE) for Rapid Mitochondrial DNA Programming of Human Cells In Vitro and Zebrafish Disease Models In Vivo.
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DOI：
10.1089/crispr.2021.0061
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发表时间：
2021-12
期刊：
影响因子：
--
通讯作者：
Ekker SC
中科院分区：
文献类型：
--
作者：
Sabharwal A;Kar B;Restrepo-Castillo S;Holmberg SR;Mathew ND;Kendall BL;Cotter RP;WareJoncas Z;Seiler C;Nakamaru-Ogiso E;Clark KJ;Ekker SC
线粒体的功能分析一直受到一些有效的方法来操纵线粒体DNA（mtDNA）和缺乏现有的动物模型。最近，TALE衍生的碱基编辑器被证明可以诱导mtDNA中的C到T（或G到A）序列变化。我们在这里报告FusX TALE碱基编辑器（FusXTBE），以促进广泛访问TALE线粒体碱基编辑技术。TALE Writer是一个从头开始的设计工具，用于绘制潜在的mtDNA碱基编辑位点。FusXTBE被证明在体外人细胞中具有与初始碱基编辑器相当的活性。斑马鱼胚胎被用作开创性的体内测试系统，FusXTBE在mtDNA基因座中诱导90+%的编辑效率，作为体内mtDNA异质性接近完全诱导的实例。对于蛋白质编码基因，观察到与单个核苷酸一样精确的基因编辑特异性。非破坏性基因分型使单动物mtDNA分析成为下游生物功能基因组应用的可能。FusXTBE是一种新的基因编辑工具包，用于探索线粒体生物学和遗传学中的重要问题。
Functional analyses of mitochondria have been hampered by few effective approaches to manipulate mitochondrial DNA (mtDNA) and a lack of existing animal models. Recently a TALE-derived base editor was shown to induce C-to-T (or G-to-A) sequence changes in mtDNA. We report here the FusX TALE Base Editor (FusXTBE) to facilitate broad-based access to TALE mitochondrial base editing technology. TALE Writer is a de novo in silico design tool to map potential mtDNA base editing sites. FusXTBE was demonstrated to function with comparable activity to the initial base editor in human cells in vitro. Zebrafish embryos were used as a pioneering in vivo test system, with FusXTBE inducing 90+% editing efficiency in mtDNA loci as an example of near-complete induction of mtDNA heteroplasmy in vivo. Gene editing specificity as precise as a single nucleotide was observed for a protein-coding gene. Nondestructive genotyping enables single-animal mtDNA analyses for downstream biological functional genomic applications. FusXTBE is a new gene editing toolkit for exploring important questions in mitochondrial biology and genetics.
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影响因子：
82.9
作者：
Gammage PA;Viscomi C;Simard ML;Costa ASH;Gaude E;Powell CA;Van Haute L;McCann BJ;Rebelo-Guiomar P;Cerutti R;Zhang L;Rebar EJ;Zeviani M;Frezza C;Stewart JB;Minczuk M
通讯作者：
Minczuk M
影响因子：
16.6
作者：
Lueck, John D.;Yoon, Jae Seok;Ahern, Christopher A.
通讯作者：
Ahern, Christopher A.
影响因子：
4.2
作者：
Ma AC;McNulty MS;Poshusta TL;Campbell JM;Martínez-Gálvez G;Argue DP;Lee HB;Urban MD;Bullard CE;Blackburn PR;Man TK;Clark KJ;Ekker SC
通讯作者：
Ekker SC
影响因子：
9.8
作者：
Hanna, MG;Nelson, IP;Wood, NW
通讯作者：
Wood, NW
DOI：
10.1042/cs20150707
发表时间：
2016-03
期刊：
Clinical science (London, England : 1979)
影响因子：
--
作者：
Rak M;Bénit P;Chrétien D;Bouchereau J;Schiff M;El-Khoury R;Tzagoloff A;Rustin P
通讯作者：
Rustin P