Bibliographic Details
| Title: |
Targeted introduction of premature stop codon in plant mitochondrial mRNA by a designer pentatricopeptide repeat protein with C‐to‐U editing function. |
| Authors: |
Manavski, Nikolay1 (AUTHOR) n.manavski@biologie.uni-muenchen.de, Abdel‐Salam, Eslam1 (AUTHOR), Schwenkert, Serena1 (AUTHOR), Kunz, Hans‐Henning2 (AUTHOR), Brachmann, Andreas3 (AUTHOR), Leister, Dario1 (AUTHOR), Meurer, Jörg1 (AUTHOR) |
| Source: |
Plant Journal. Feb2025, Vol. 121 Issue 3, p1-12. 12p. |
| Subject Terms: |
*RNA editing, *PENTATRICOPEPTIDE repeat genes, *GENE expression, *STOP codons, *ARABIDOPSIS thaliana, *NADH dehydrogenase, *MITOCHONDRIAL RNA |
| Abstract: |
SUMMARY: RNA editing is a crucial post‐transcriptional modification in endosymbiotic plant organelles, predominantly involving C‐to‐U conversions. Pentatricopeptide repeat (PPR) proteins play a key role in this process. To establish a system for gene expression manipulation in genetically inaccessible mitochondria, we engineered a synthetic PPR protein, dPPR‐nad7‐DYW, to induce de novo C‐to‐U editing in the NADH dehydrogenase subunit 7 (nad7) mRNA of Arabidopsis thaliana, thereby creating a premature stop codon. This designer protein, composed of 13 P‐type PPR domains, was fused with the DYW‐type cytidine deaminase domain from Physcomitrium patens PpPPR_56 and programmed to bind a specific nad7 mRNA segment. In vitro binding assays confirmed the specificity of dPPR‐nad7‐DYW for its target sequence. When expressed in Arabidopsis plants, dPPR‐nad7‐DYW achieved up to 85% editing efficiency at the target site, successfully introducing a premature stop codon in nad7 mRNA. This resulted in reduced polysome loading of nad7 transcripts and a phenotype characteristic of mitochondrial complex I dysfunction. RNA‐sequencing revealed potential off‐target editing events, albeit at lower frequencies. Our study demonstrates the successful application of an editing factor with a synthetic P‐type PPR tract targeting a de novo editing site in plant mitochondria, achieving high editing efficiency. This approach opens new avenues for manipulating organellar gene expression and studying mitochondrial gene function in plants and other eukaryotes. [ABSTRACT FROM AUTHOR] |
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| Database: |
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