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RNA Synthesis Using the CEM Group

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Abstract

We have developed a solid-phase synthesis of RNA oligomers with 2-cyanoethoxymethyl (CEM) as the 2′-hydroxyl protecting group. The method allows the synthesis of RNA oligomers with high efficiency and high purity. In this section, we describe the synthesis of CEM amidites and RNA synthesis by using CEM amidites. The advantages of the CEM group include its low steric hindrance, leading to a high coupling yield, and its ease of removal under mild conditions without any decomposition of the RNA oligomers.

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References

  1. Usman N, Ogilvie KK, Jiang M-Y, Cedergren RJ (1987) The automated chemical synthesis of long oligoribonucleotides using 2′-O-silylated ribonucleotides 3′-O-phosphoramidites on a controlled-pore glass support: synthesis of a 43-nucleotide sequence similar to the 3′-half molecule of an Escherichia coli formylmethionine tRNA. J Am Chem Soc 109:7845–7854

    Article  CAS  Google Scholar 

  2. Gough GR, Miller TJ, Mantick NA (1996) p-Nitrobenzyloxymethyl: a new fluoride-removable protecting group for ribonucleoside 2′-hydroxyls. Tetrahedron Lett 37:981–982

    Article  CAS  Google Scholar 

  3. Welz R, Müller S (2002) 5-(benzylmercapto)-1H-tetrazole as activator for 2′-O-TBDMS phosphoramidite building blocks in RNA synthesis. Tetrahedron Lett 43:795–797

    Article  CAS  Google Scholar 

  4. Scaringe SA, Wincott FE, Caruthers MH (1998) Novel RNA synthesis method using 5′-O-silyl-2′-O-orthoester protecting groups. J Am Chem Soc 120:11820–11821

    Article  CAS  Google Scholar 

  5. Pitsch S, Weiss PA, Jenny L, Stutz A, Wu X (2001) Reliable chemical synthesis of oligoribonucleotides (RNA) with 2′-O-[(triisopropylsilyl)oxy]methyl (2′-O-tom)-protected phosphoramidites. Helv Chim Acta 84:3773–3795

    Article  CAS  Google Scholar 

  6. Semenyuk A, Földesi A, Johansson T, Estmer-Nilsson C, Blomgren P, Brännvall M, Kirsebom LA, Kwiatkowski M (2006) Synthesis of RNA using 2′-O-DTM protection. J Am Chem Soc 128:12356–12357

    Article  CAS  PubMed  Google Scholar 

  7. Micura R (2002) Small interfering RNAs and their chemical synthesis. Angew Chem Int Ed 41:2265–2269

    Article  CAS  Google Scholar 

  8. Ohgi T, Masutomi Y, Ishiyama K, Kitagawa H, Shiba Y, Yano J (2005) A new RNA synthetic method with a 2′-O-(2-cyanoethoxymethyl) protecting group (CEM). Org Lett 7:3477–3480

    Article  CAS  PubMed  Google Scholar 

  9. Shiba Y, Masuda H, Watanabe N, Ego T, Takagaki K, Ishiyama K, Ohgi T, Yano J (2007) Chemical synthesis of a very long oligoribonucleotide with 2-cyanoethoxymethyl (CEM) as the 2′-O-protecting group: structural identification and biological activity of a synthetic 110 mer precursor-microRNA candidate. Nucleic Acid Res 35:3287–3296

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Nagata S, Hamasaki T, Uetake K, Masuda H, Takagaki K, Oka N, Wada T, Ohgi T, Yano J (2010) Synthesis and biological activity of artificial mRNA prepared with novel phosphorylating reagents. Nucleic Acid Res 38:7845–7857

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

The research described here was supported in part by grants from the New Energy and Industrial Technology Development Organization (NEDO) of Japan for its Functional RNA Project. We thank Dr. G. E. Smyth, Discovery Research Laboratories in Tsukuba, Nippon Shinyaku Co., for helpful discussions and suggestions concerning the manuscript.

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Correspondence to Hidetoshi Kitagawa .

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Kitagawa, H. (2018). RNA Synthesis Using the CEM Group. In: Obika, S., Sekine, M. (eds) Synthesis of Therapeutic Oligonucleotides. Springer, Singapore. https://doi.org/10.1007/978-981-13-1912-9_4

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