PUBLICATION
N1-methylpseudouridine mRNA modification enhances efficiency and specificity of gene overexpression by preventing Prkra-mediated global translation repression
- Authors
- Lu, T., Chen, A., Li, C., Li, K., Wang, S., Zhang, Y., Yang, B., Wang, J., Gong, Q., Li, A., Liu, X., Ma, P., Mao, B., Shi, D.L., Shao, M.
- ID
- ZDB-PUB-251020-4
- Date
- 2025
- Source
- Nucleic acids research 53: (Journal)
- Registered Authors
- Shao, Ming
- Keywords
- none
- MeSH Terms
-
- Animals
- Embryo, Nonmammalian/metabolism
- Protein Biosynthesis*
- Pseudouridine*/analogs & derivatives
- Pseudouridine*/metabolism
- RNA, Double-Stranded/genetics
- RNA, Double-Stranded/metabolism
- RNA, Messenger*/chemistry
- RNA, Messenger*/genetics
- RNA, Messenger*/metabolism
- Zebrafish/embryology
- Zebrafish/genetics
- Zebrafish Proteins*/genetics
- Zebrafish Proteins*/metabolism
- PubMed
- 41099713 Full text @ Nucleic Acids Res.
Citation
Lu, T., Chen, A., Li, C., Li, K., Wang, S., Zhang, Y., Yang, B., Wang, J., Gong, Q., Li, A., Liu, X., Ma, P., Mao, B., Shi, D.L., Shao, M. (2025) N1-methylpseudouridine mRNA modification enhances efficiency and specificity of gene overexpression by preventing Prkra-mediated global translation repression. Nucleic acids research. 53:.
Abstract
In vitro transcribed messenger RNA (IVT mRNA) has emerged as a pivotal tool in mRNA-based therapies and has been extensively employed in gene function studies and genetic tool applications. However, the IVT process generates double-stranded RNA (dsRNA) by-products that are recognized by dsRNA sensors, triggering innate immune responses. In this study, we comprehensively analyzed the detrimental effects of dsRNA by-products on early zebrafish embryos, revealing that these by-products induce cell necrosis and delay maternal-zygotic transition (MZT) by reducing global translation efficiency via Prkra (Protein Activator Of Interferon Induced Protein Kinase; also called PACT in mammals), a dsRNA sensor recently identified in pluripotent cells. Importantly, we demonstrate that N1-methylpseudouridine (m1Ψ) modification of IVT mRNAs effectively mitigates these adverse effects, as m1Ψ-modified dsRNAs exhibit significantly lower binding affinity to the Prkra dimer. Our findings underscore a previously overlooked challenge in the use of IVT mRNA in early embryos and offer a robust solution to enhance the fidelity of mRNA applications. Furthermore, we elucidate that m1Ψ modification minimizes the dsRNA-induced stress response in pluripotent cells through a distinct mechanism.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping