TY - JOUR
T1 - Complete Chemical Synthesis of Minimal Messenger RNA by Efficient Chemical Capping Reaction
AU - Abe, Naoko
AU - Imaeda, Akihiro
AU - Inagaki, Masahito
AU - Li, Zhenmin
AU - Kawaguchi, Daisuke
AU - Onda, Kaoru
AU - Nakashima, Yuko
AU - Uchida, Satoshi
AU - Hashiya, Fumitaka
AU - Kimura, Yasuaki
AU - Abe, Hiroshi
N1 - Funding Information:
This work was supported by the Japan Society for the Promotion of Science (JSPS) [Grant-in-Aid for Scientific Research (B) 16H04178 to H.A.; Grant-in-Aid for Early-Career Scientists 18K14357 to N.A.], the Japan Science and Technology Agency (JST) [CREST JPMJCR18S1 to H.A.), and the Japan Agency for Medical Research and Development (AMED) [LEAP JP21gm0010008 to H.A.]. We thank the Chemical Instrumentation Facility of the Research Center for Materials Science at Nagoya University for allowing us to conduct the analyses. We are grateful to R. Ogisu for technical assistance with the RNA synthesis.
Publisher Copyright:
© 2022 American Chemical Society.
PY - 2021
Y1 - 2021
N2 - Site-specific chemical modification of mRNA can improve its translational efficiency and stability. For this purpose, it is desirable to develop a complete chemical synthesis method for chemically modified mRNA. The key is a chemical reaction that introduces a cap structure into the chemically synthesized RNA. In this study, we developed a fast and quantitative chemical capping reaction between 5′-phosphorylated RNA and N7-methylated GDP imidazolide in the presence of 1-methylimidazole in the organic solvent dimethyl sulfoxide. It enabled quantitative preparation of capping RNA within 3 h. We prepared chemically modified 107-nucleotide mRNAs, including N6-methyladenosine, insertion of non-nucleotide linkers, and 2′-O-methylated nucleotides at the 5′ end and evaluated their effects on translational activity in cultured HeLa cells. The results showed that mRNAs with non-nucleotide linkers in the untranslated regions were sufficiently tolerant to translation and that mRNAs with the Cap_2 structure had higher translational activity than those with the Cap_0 structure.
AB - Site-specific chemical modification of mRNA can improve its translational efficiency and stability. For this purpose, it is desirable to develop a complete chemical synthesis method for chemically modified mRNA. The key is a chemical reaction that introduces a cap structure into the chemically synthesized RNA. In this study, we developed a fast and quantitative chemical capping reaction between 5′-phosphorylated RNA and N7-methylated GDP imidazolide in the presence of 1-methylimidazole in the organic solvent dimethyl sulfoxide. It enabled quantitative preparation of capping RNA within 3 h. We prepared chemically modified 107-nucleotide mRNAs, including N6-methyladenosine, insertion of non-nucleotide linkers, and 2′-O-methylated nucleotides at the 5′ end and evaluated their effects on translational activity in cultured HeLa cells. The results showed that mRNAs with non-nucleotide linkers in the untranslated regions were sufficiently tolerant to translation and that mRNAs with the Cap_2 structure had higher translational activity than those with the Cap_0 structure.
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U2 - 10.1021/acschembio.1c00996
DO - 10.1021/acschembio.1c00996
M3 - Article
C2 - 35608277
AN - SCOPUS:85131857396
SN - 1554-8929
JO - ACS Chemical Biology
JF - ACS Chemical Biology
ER -