biostudies-literatureUnknown27Valentini PSINEUPs are a novel class of natural and synthetic non-coding antisense RNA molecules able to increase the translation of a target mRNA. They present a modular organization comprising an unstructured antisense target-specific domain, which sets the specificity of each individual SINEUP, and a structured effector domain, which is responsible for the translation enhancement. In order to design a fully functional <i>in vitro</i> transcribed SINEUP for therapeutics applications, SINEUP RNAs were synthesized <i>in vitro</i> with a variety of chemical modifications and screened for their activity on endogenous target mRNA upon transfection. Three combinations of modified ribonucleotides-2'O methyl-ATP (Am), N6 methyl-ATP (m6A), and pseudo-UTP (ψ)-conferred SINEUP activity to naked RNA. The best Molecular therapy. Nucleic acids1092-1102https://www.ebi.ac.uk/biostudies/studies/S-EPMC8857549biostudies-literatureTowards SINEUP-based therapeutics: Design of an <i>in vitro</i> synthesized SINEUP RNA.PMC8857549Valentini PZacco EMangoni DCarninci PEspinoza SAndrews BWebster NAGustincich SPandolfini LPierattini BTartaglia GGfalseTowards SINEUP-based therapeutics: Design of an <i>in vitro</i> synthesized SINEUP RNA.SINEUPs are a novel class of natural and synthetic non-coding antisense RNA molecules able to increase the translation of a target mRNA. They present a modular organization comprising an unstructured antisense target-specific domain, which sets the specificity of each individual SINEUP, and a structured effector domain, which is responsible for the translation enhancement. In order to design a fully functional <i>in vitro</i> transcribed SINEUP for therapeutics applications, SINEUP RNAs were synthesized <i>in vitro</i> with a variety of chemical modifications and screened for their activity on endogenous target mRNA upon transfection. Three combinations of modified ribonucleotides-2'O methyl-ATP (Am), N6 methyl-ATP (m6A), and pseudo-UTP (ψ)-conferred SINEUP activity to naked RNA. The best 2022-01-01T00:00:00Z2022 Mar2025-04-22T04:44:53.288Z2025-04-05T21:02:54.971ZS-EPMC88575493522890210.1016/j.omtn.2022.01.021