Targeted introduction of premature stop codon in plant mitochondrial mRNA by a designer pentatricopeptide repeat protein with C-to-U editing function.

ABSTRACT: 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.

SUBMITTER: Manavski N 

PROVIDER: S-EPMC11803495 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature