2. Academic Validation
  3. Cap analogs with a hydrophobic photocleavable tag enable facile purification of fully capped mRNA with various cap structures

Cap analogs with a hydrophobic photocleavable tag enable facile purification of fully capped mRNA with various cap structures

  • Nat Commun. 2023 May 11;14(1):2657. doi: 10.1038/s41467-023-38244-8.
Masahito Inagaki # 1,  Naoko Abe # 1,  Zhenmin Li 1,  Yuko Nakashima 1 2,  Susit Acharyya 1,  Kazuya Ogawa 1,  Daisuke Kawaguchi 1,  Haruka Hiraoka 1,  Ayaka Banno 1,  Zheyu Meng 1,  Mizuki Tada 1,  Tatsuma Ishida 1,  Pingxue Lyu 1,  Kengo Kokubo 1,  Hirotaka Murase 1,  Fumitaka Hashiya 2,  Yasuaki Kimura 1,  Satoshi Uchida 3 4 5,  Hiroshi Abe 6 7 8 9
Affiliations

Affiliations

  • 1 Department of Chemistry, Graduate School of Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi, 464-8602, Japan.
  • 2 Research Center for Materials Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi, 464-8602, Japan.
  • 3 Graduate School of Medical Science, Kyoto Prefectural University of Medicine, 1-5 Shimogamohangi-cho, Sakyo-ku, Kyoto, 606-0823, Japan.
  • 4 Innovation Center of NanoMedicine (iCONM), Kawasaki Institute of Industrial Promotion, 3-25-14 Tonomachi, Kawasaki-ku, Kawasaki, 210-0821, Japan.
  • 5 Medical Research Institute, Tokyo Medical and Dental University (TMDU), 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8510, Japan.
  • 6 Department of Chemistry, Graduate School of Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi, 464-8602, Japan. h-abe@chem.nagoya-u.ac.jp.
  • 7 Research Center for Materials Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi, 464-8602, Japan. h-abe@chem.nagoya-u.ac.jp.
  • 8 CREST, Japan Science and Technology Agency, 7, Gobancho, Chiyoda-ku, Tokyo, 102-0076, Japan. h-abe@chem.nagoya-u.ac.jp.
  • 9 Institute for Glyco-core Research (iGCORE), Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi, 464-8601, Japan. h-abe@chem.nagoya-u.ac.jp.
  • # Contributed equally.
PMID: 37169757 DOI: 10.1038/s41467-023-38244-8
Abstract

Starting with the clinical application of two vaccines in 2020, mRNA therapeutics are currently being investigated for a variety of applications. Removing immunogenic uncapped mRNA from transcribed mRNA is critical in mRNA research and clinical applications. Commonly used capping methods provide maximum capping efficiency of around 80-90% for widely used Cap-0- and Cap-1-type mRNAs. However, uncapped and capped mRNA possesses almost identical physicochemical properties, posing challenges to their physical separation. In this work, we develop hydrophobic photocaged tag-modified Cap Analogs, which separate capped mRNA from uncapped mRNA by reversed-phase high-performance liquid chromatography. Subsequent photo-irradiation recovers footprint-free native capped mRNA. This approach provides 100% capping efficiency even in Cap-2-type mRNA with versatility applicable to 650 nt and 4,247 nt mRNA. We find that the Cap-2-type mRNA shows up to 3- to 4-fold higher translation activity in cultured cells and Animals than the Cap-1-type mRNA prepared by the standard capping method.

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