Site-Specific Mitochondrial RNA N1-Methyladenosine Demethylation via an Engineered MTS-PUF-ALKBH3 Fusion Protein.

ABSTRACT: Mitochondrial RNA N1-methyladenosine (m¹A) is a prevalent and reversible epitranscriptomic modification. While the biological roles of cytosolic m¹A have been increasingly understood, the causal relationship between site-specific mitochondrial m¹A and phenotypic outcomes remain elusive, partly due to the lack of precise editing tools. Here, a CRISPR-free mitochondrial RNA m¹A demethylation (MRD) editor is reported, which fuses mitochondria-localized engineered PUF RNA-binding protein with the m¹A demethylase ALKBH3. Independent cellular assays across multiple sites confirm that MRD editor enables precise demethylation of m¹A in mitochondrial mRNAs and tRNAs, leading to correlated changes in mitochondrial protein levels with

SUBMITTER: Li X 

PROVIDER: S-EPMC12767104 | biostudies-literature | 2026 Jan

REPOSITORIES: biostudies-literature