Protein coding mitochondrial-targeted RNAs rescue mitochondrial disease in vivo.
Protein coding mitochondrial-targeted RNAs rescue mitochondrial disease in vivo.
复制标题
编码线粒体靶向 RNA 的蛋白质可在体内挽救线粒体疾病。
DOI：
10.1016/j.nbd.2018.06.009
复制
发表时间：
2018
影响因子：
6.1
通讯作者：
Palladino,MichaelJ
中科院分区：
文献类型：
--
作者：
Markantone,DesireeM;Towheed,Atif;Crain,AaronT;Collins,JessicaM;Celotto,AliciaM;Palladino,MichaelJ
线粒体脑肌病（ME）由氧化磷酸化关键线粒体基因突变引起。严重和无法治疗的ME是由影响每个内源性线粒体编码基因的突变引起的，包括所有13个已建立的蛋白质编码基因。操纵线粒体基因组的有效技术是有限的，并且靶向线粒体蛋白表达目前不可用。在这里，我们报告的发展amitochondrial-targeted RNA表达（mtTRES）载体能够在线粒体内的蛋白质表达（mtTRESPro）。我们证明了mtTRESPro表达的RNA靶向于线粒体，并且能够使用EGFP编码的构建体在体内翻译。我们还检测了编码野生型ATP 6的mtTRESProconstructures拯救已建立的ATP 61果蝇ME模型的能力。检测了遗传拯救，包括与线粒体靶向翻译相关物TLI-NCL：：ATP 6 RNA共表达的检测，其功能是降低内源性突变蛋白的表达。数据表明，线粒体靶向野生型ATP 6编码RNA的异位RNA表达足以部分挽救严重的和建立的ME动物模型，但仅当与抑制突变蛋白表达的方法结合时，突变蛋白可能竞争掺入复合物V中。
Mitochondrial encephalomyopathies (MEs) result from mutations in mitochondrial genes critical to oxidative phosphorylation. Severe and untreatable ME results from mutations affecting each endogenous mitochondrial encoded gene, including all 13 established protein coding genes. Effective techniques to manipulate mitochondrial genome are limited and targeted mitochondrial protein expression is currently unavailable. Here we report the development of amitochondrial-targeted RNA expression(mtTRES) vector capable of protein expression within mitochondria (mtTRESPro). We demonstrate thatmtTRESProexpressed RNAs are targeted to mitochondria and are capable of being translated using EGFP encoded constructsin vivo. We additionally testmtTRESProconstructs encoding wild type ATP6 for their ability to rescue an establishedATP61Drosophilamodel of ME. Genetic rescue is examined including tests with co-expression of mitochondrial targeted translational inhibitorsTLI-NCL::ATP6RNAs that function to reduce expression of the endogenous mutant protein. The data demonstrate allotopic RNA expression of mitochondrial targeted wild typeATP6coding RNAs are sufficient to partially rescue a severe and established animal model of ME but only when combined with a method to inhibit mutant protein expression, which likely competes for incorporation into complex V.