RNA-Based Translation Activators for Targeted Gene Upregulation

Authors

Yang Cao
Huachun Liu
Shannon S Lu
Krysten A Jones
Anitha P Govind
Okunola Jeyifous
Christine Q Simmons
Negar Tabatabaei
William N Green
Jimmy L Holder
Soroush Tahmasebi
Alfred L George
Bryan C Dickinson

Publication Date

10-26-2023

Journal

Nature Communications

DOI

10.1038/s41467-023-42252-z

PMID

37884512

PMCID

PMC10603104

PubMedCentral® Posted Date

10-26-2023

PubMedCentral® Full Text Version

Post-print

Published Open-Access

yes

Keywords

Animals, Mice, Humans, Protein Biosynthesis, Up-Regulation, Gene Expression Regulation, RNA, Messenger, Internal Ribosome Entry Sites, Mammals, Synthetic biology, RNA, Translation, Ribosome, Gene regulation

Abstract

Technologies capable of programmable translation activation offer strategies to develop therapeutics for diseases caused by insufficient gene expression. Here, we present "translation-activating RNAs" (taRNAs), a bifunctional RNA-based molecular technology that binds to a specific mRNA of interest and directly upregulates its translation. taRNAs are constructed from a variety of viral or mammalian RNA internal ribosome entry sites (IRESs) and upregulate translation for a suite of target mRNAs. We minimize the taRNA scaffold to 94 nucleotides, identify two translation initiation factor proteins responsible for taRNA activity, and validate the technology by amplifying SYNGAP1 expression, a haploinsufficiency disease target, in patient-derived cells. Finally, taRNAs are suitable for delivery as RNA molecules by lipid nanoparticles (LNPs) to cell lines, primary neurons, and mouse liver in vivo. taRNAs provide a general and compact nucleic acid-based technology to upregulate protein production from endogenous mRNAs, and may open up possibilities for therapeutic RNA research.