SINEUP non-coding RNA activity depends on specific N6-methyladenosine nucleotides
SINEUPs are natural and synthetic antisense long non-coding RNAs (lncRNAs) selectively enhancing target mRNAs translation by increasing their association with polysomes. This activity requires two RNA domains: an embedded inverted SINEB2 element acting as effector domain, and an antisense region, th...
| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-06-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253123000884 |
| _version_ | 1797839040905478144 |
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| author | Bianca Pierattini Sabrina D’Agostino Carlotta Bon Omar Peruzzo Andrej Alendar Azzurra Codino Gloria Ros Francesca Persichetti Remo Sanges Piero Carninci Claudio Santoro Stefano Espinoza Paola Valentini Luca Pandolfini Stefano Gustincich |
| author_facet | Bianca Pierattini Sabrina D’Agostino Carlotta Bon Omar Peruzzo Andrej Alendar Azzurra Codino Gloria Ros Francesca Persichetti Remo Sanges Piero Carninci Claudio Santoro Stefano Espinoza Paola Valentini Luca Pandolfini Stefano Gustincich |
| author_sort | Bianca Pierattini |
| collection | DOAJ |
| description | SINEUPs are natural and synthetic antisense long non-coding RNAs (lncRNAs) selectively enhancing target mRNAs translation by increasing their association with polysomes. This activity requires two RNA domains: an embedded inverted SINEB2 element acting as effector domain, and an antisense region, the binding domain, conferring target selectivity. SINEUP technology presents several advantages to treat genetic (haploinsufficiencies) and complex diseases restoring the physiological activity of diseased genes and of compensatory pathways. To streamline these applications to the clinic, a better understanding of the mechanism of action is needed. Here we show that natural mouse SINEUP AS Uchl1 and synthetic human miniSINEUP-DJ-1 are N6-methyladenosine (m6A) modified by METTL3 enzyme. Then, we map m6A-modified sites along SINEUP sequence with Nanopore direct RNA sequencing and a reverse transcription assay. We report that m6A removal from SINEUP RNA causes the depletion of endogenous target mRNA from actively translating polysomes, without altering SINEUP enrichment in ribosomal subunit-associated fractions. These results prove that SINEUP activity requires an m6A-dependent step to enhance translation of target mRNAs, providing a new mechanism for m6A translation regulation and strengthening our knowledge of SINEUP-specific mode of action. Altogether these new findings pave the way to a more effective therapeutic application of this well-defined class of lncRNAs. |
| first_indexed | 2024-04-09T15:51:50Z |
| format | Article |
| id | doaj.art-439e7dc3410e4a769903132bc7c5d494 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-04-09T15:51:50Z |
| publishDate | 2023-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-439e7dc3410e4a769903132bc7c5d4942023-04-26T05:58:24ZengElsevierMolecular Therapy: Nucleic Acids2162-25312023-06-0132402414SINEUP non-coding RNA activity depends on specific N6-methyladenosine nucleotidesBianca Pierattini0Sabrina D’Agostino1Carlotta Bon2Omar Peruzzo3Andrej Alendar4Azzurra Codino5Gloria Ros6Francesca Persichetti7Remo Sanges8Piero Carninci9Claudio Santoro10Stefano Espinoza11Paola Valentini12Luca Pandolfini13Stefano Gustincich14Area of Neuroscience, International School for Advanced Studies (SISSA), Trieste, Italy; Central RNA Laboratory, Istituto Italiano di Tecnologia (IIT), Genova, ItalyCentral RNA Laboratory, Istituto Italiano di Tecnologia (IIT), Genova, ItalyCentral RNA Laboratory, Istituto Italiano di Tecnologia (IIT), Genova, ItalyCentral RNA Laboratory, Istituto Italiano di Tecnologia (IIT), Genova, ItalyThe Gurdon Institute and Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UKCentral RNA Laboratory, Istituto Italiano di Tecnologia (IIT), Genova, ItalyDepartment of Health Sciences and Research Center on Autoimmune and Allergic Diseases (CAAD), University of Piemonte Orientale (UPO), Novara, ItalyDepartment of Health Sciences and Research Center on Autoimmune and Allergic Diseases (CAAD), University of Piemonte Orientale (UPO), Novara, ItalyArea of Neuroscience, International School for Advanced Studies (SISSA), Trieste, Italy; Central RNA Laboratory, Istituto Italiano di Tecnologia (IIT), Genova, ItalyRIKEN Center for Integrative Medical Sciences (IMS), Yokohama 230-0045, Japan; Human Technopole, 20157 Milan, ItalyDepartment of Health Sciences and Research Center on Autoimmune and Allergic Diseases (CAAD), University of Piemonte Orientale (UPO), Novara, ItalyCentral RNA Laboratory, Istituto Italiano di Tecnologia (IIT), Genova, Italy; Department of Health Sciences and Research Center on Autoimmune and Allergic Diseases (CAAD), University of Piemonte Orientale (UPO), Novara, ItalyCentral RNA Laboratory, Istituto Italiano di Tecnologia (IIT), Genova, Italy; Corresponding author: Paola Valentini, Central RNA Laboratory, Istituto Italiano di Tecnologia (IIT), via Melen 83, 16152 Genova, Italy.Central RNA Laboratory, Istituto Italiano di Tecnologia (IIT), Genova, Italy; Corresponding author: Luca Pandolfini, Central RNA Laboratory, Istituto Italiano di Tecnologia (IIT), via Melen 83, 16152 Genova, Italy.Central RNA Laboratory, Istituto Italiano di Tecnologia (IIT), Genova, Italy; Corresponding author: Stefano Gustincich, Central RNA Laboratory, Istituto Italiano di Tecnologia (IIT), via Melen 83, 16152 Genova, Italy.SINEUPs are natural and synthetic antisense long non-coding RNAs (lncRNAs) selectively enhancing target mRNAs translation by increasing their association with polysomes. This activity requires two RNA domains: an embedded inverted SINEB2 element acting as effector domain, and an antisense region, the binding domain, conferring target selectivity. SINEUP technology presents several advantages to treat genetic (haploinsufficiencies) and complex diseases restoring the physiological activity of diseased genes and of compensatory pathways. To streamline these applications to the clinic, a better understanding of the mechanism of action is needed. Here we show that natural mouse SINEUP AS Uchl1 and synthetic human miniSINEUP-DJ-1 are N6-methyladenosine (m6A) modified by METTL3 enzyme. Then, we map m6A-modified sites along SINEUP sequence with Nanopore direct RNA sequencing and a reverse transcription assay. We report that m6A removal from SINEUP RNA causes the depletion of endogenous target mRNA from actively translating polysomes, without altering SINEUP enrichment in ribosomal subunit-associated fractions. These results prove that SINEUP activity requires an m6A-dependent step to enhance translation of target mRNAs, providing a new mechanism for m6A translation regulation and strengthening our knowledge of SINEUP-specific mode of action. Altogether these new findings pave the way to a more effective therapeutic application of this well-defined class of lncRNAs.http://www.sciencedirect.com/science/article/pii/S2162253123000884MT: Non-coding RNAsSINEUPlncRNARNA therapeuticsm6AN6-methyladenosine |
| spellingShingle | Bianca Pierattini Sabrina D’Agostino Carlotta Bon Omar Peruzzo Andrej Alendar Azzurra Codino Gloria Ros Francesca Persichetti Remo Sanges Piero Carninci Claudio Santoro Stefano Espinoza Paola Valentini Luca Pandolfini Stefano Gustincich SINEUP non-coding RNA activity depends on specific N6-methyladenosine nucleotides Molecular Therapy: Nucleic Acids MT: Non-coding RNAs SINEUP lncRNA RNA therapeutics m6A N6-methyladenosine |
| title | SINEUP non-coding RNA activity depends on specific N6-methyladenosine nucleotides |
| title_full | SINEUP non-coding RNA activity depends on specific N6-methyladenosine nucleotides |
| title_fullStr | SINEUP non-coding RNA activity depends on specific N6-methyladenosine nucleotides |
| title_full_unstemmed | SINEUP non-coding RNA activity depends on specific N6-methyladenosine nucleotides |
| title_short | SINEUP non-coding RNA activity depends on specific N6-methyladenosine nucleotides |
| title_sort | sineup non coding rna activity depends on specific n6 methyladenosine nucleotides |
| topic | MT: Non-coding RNAs SINEUP lncRNA RNA therapeutics m6A N6-methyladenosine |
| url | http://www.sciencedirect.com/science/article/pii/S2162253123000884 |
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