Chemical and topological design of multicapped mRNA and capped circular RNA to augment translation

Hongyu Chen; Dangliang Liu; Abhishek Aditham; Jianting Guo; Jiahao Huang; Franklin Kostas; Kamal Maher; Mirco J Friedrich; Ramnik J Xavier; Feng Zhang; Xiao Wang

doi:10.1038/s41587-024-02393-y

Chemical and topological design of multicapped mRNA and capped circular RNA to augment translation

Nat Biotechnol. 2024 Sep 23. doi: 10.1038/s41587-024-02393-y. Online ahead of print.

Authors

Hongyu Chen^#^{1

2}, Dangliang Liu^#^{1

2}, Abhishek Aditham^#^{2

3}, Jianting Guo^{1

2}, Jiahao Huang^{1

2}, Franklin Kostas^{1

2}, Kamal Maher^{2

4}, Mirco J Friedrich^{2

3

5

6

7}, Ramnik J Xavier^{2

8}, Feng Zhang^{2

3

5

6

7}, Xiao Wang^{9

10

11}

Affiliations

¹ Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, USA.
² Broad Institute of MIT and Harvard, Cambridge, MA, USA.
³ Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
⁴ Computational and Systems Biology Program, Massachusetts Institute of Technology, Cambridge, MA, USA.
⁵ Howard Hughes Medical Institute, Cambridge, MA, USA.
⁶ McGovern Institute for Brain Research at MIT, Cambridge, MA, USA.
⁷ Department of Brain and Cognitive Science, Massachusetts Institute of Technology, Cambridge, MA, USA.
⁸ Center for Computational and Integrative Biology, Department of Molecular Biology, Massachusetts General Hospital, Boston, MA, USA.
⁹ Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA, USA. [email protected].
¹⁰ Broad Institute of MIT and Harvard, Cambridge, MA, USA. [email protected].
¹¹ Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA, USA. [email protected].

^# Contributed equally.

PMID: 39313647
DOI: 10.1038/s41587-024-02393-y

Abstract

Protein and vaccine therapies based on mRNA would benefit from an increase in translation capacity. Here, we report a method to augment translation named ligation-enabled mRNA-oligonucleotide assembly (LEGO). We systematically screen different chemotopological motifs and find that a branched mRNA cap effectively initiates translation on linear or circular mRNAs without internal ribosome entry sites. Two types of chemical modification, locked nucleic acid (LNA) N⁷-methylguanosine modifications on the cap and LNA + 5 × 2' O-methyl on the 5' untranslated region, enhance RNA-eukaryotic translation initiation factor (eIF4E-eIF4G) binding and RNA stability against decapping in vitro. Through multidimensional chemotopological engineering of dual-capped mRNA and capped circular RNA, we enhanced mRNA protein production by up to tenfold in vivo, resulting in 17-fold and 3.7-fold higher antibody production after prime and boost doses in a severe acute respiratory syndrome coronavirus 2 vaccine setting, respectively. The LEGO platform opens possibilities to design unnatural RNA structures and topologies beyond canonical linear and circular RNAs for both basic research and therapeutic applications.

Abstract

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