Mutations in the small genome present in mitochondria often result in severe pathologies.Different genetic strategies have been explored,aiming to rescue such mutations.A number of these strategies were based on the c...Mutations in the small genome present in mitochondria often result in severe pathologies.Different genetic strategies have been explored,aiming to rescue such mutations.A number of these strategies were based on the capacity of human mitochondria to import RNAs from the cytosol and designed to repress the replication of the mutated genomes or to provide the organelles with wild-type versions of mutant transcripts.However,the mutant RNAs present in mitochondria turned out to be an obstacle to therapy and little attention has been devoted so far to their elimination.Here,we present the development of a strategy to knockdown mitochondrial RNAs in human cells using the transfer RNA-like structure of Brome mosaic virus or Tobacco mosaic virus as a shuttle to drive trans-cleaving ribozymes into the organelles in human cell lines.We obtained a specific knockdown of the targeted mitochondrial ATP6 mRNA,followed by a deep drop in ATP6 protein and a functional impairment of the oxidative phosphorylation chain.Our strategy provides a powerful approach to eliminate mutant organellar transcripts and to analyse the control and communication of the human organellar genetic system.展开更多
基金supported by a grant from the Polish Medical Research Agency(2021/ABM/05/00004)The research was further funded by grants from the Polish National Science Centre(2016/21/N/NZ1/00564)+2 种基金the French State Program‘Investments for the future’(LABEX ANR-11-LABX-0057_MITOCROSS and ANR-10-LABX-0040-SPS)the French National Research Agency(ANR-06-MRAR-037-02 and ANR-09-BLAN-0240-01)R.V.was supported by a fellowship from CNRS and the French Région Alsace.
文摘Mutations in the small genome present in mitochondria often result in severe pathologies.Different genetic strategies have been explored,aiming to rescue such mutations.A number of these strategies were based on the capacity of human mitochondria to import RNAs from the cytosol and designed to repress the replication of the mutated genomes or to provide the organelles with wild-type versions of mutant transcripts.However,the mutant RNAs present in mitochondria turned out to be an obstacle to therapy and little attention has been devoted so far to their elimination.Here,we present the development of a strategy to knockdown mitochondrial RNAs in human cells using the transfer RNA-like structure of Brome mosaic virus or Tobacco mosaic virus as a shuttle to drive trans-cleaving ribozymes into the organelles in human cell lines.We obtained a specific knockdown of the targeted mitochondrial ATP6 mRNA,followed by a deep drop in ATP6 protein and a functional impairment of the oxidative phosphorylation chain.Our strategy provides a powerful approach to eliminate mutant organellar transcripts and to analyse the control and communication of the human organellar genetic system.
