Chemical synthesis of a very long RNA oligomer, a 110mer precursor-miRNA candidate, with 2-cyanoethoxymethyl (CEM) as the 2'-O-protecting group | Zendy

Hirotoshi MASUDA | Zendy; Yoshiaki Shiba | Zendy; Naoki Watanabe | Zendy; Kazuchika Takagaki | Zendy; Koichi Ishiyama | Zendy; Tadaaki Ohgi | Zendy; JunIchi Yano | Zendy

Open Access

Chemical synthesis of a very long RNA oligomer, a 110mer precursor-miRNA candidate, with 2-cyanoethoxymethyl (CEM) as the 2'-O-protecting group

Author(s) -

Hirotoshi MASUDA,

Yoshiaki Shiba,

Naoki Watanabe,

Kazuchika Takagaki,

Koichi Ishiyama,

Tadaaki Ohgi,

JunIchi Yano

Publication year - 2007

Publication title -

nucleic acids symposium series

Language(s) - English

Resource type - Journals

eISSN - 1746-8272

pISSN - 0261-3166

DOI - 10.1093/nass/nrm002

Subject(s) - oligomer , rna , rnase p , phosphoramidite , oligonucleotide , chemistry , combinatorial chemistry , nucleotide , chemical synthesis , microrna , mass spectrometry , protecting group , biochemistry , in vitro , gene , organic chemistry , chromatography , alkyl

A long RNA oligomer, a 110mer with the sequence of a precursor-miRNA candidate, has been chemically synthesized in a single synthesizer run by means of standard automated phosphoramidite chemistry. The synthetic method involved the use of 2-cyanoethoxymethyl (CEM), a 2'-hydroxyl protecting group recently developed in our laboratory. We confirmed the identity of the synthetic 110mer by MALDI-TOF mass spectrometry, as well as HPLC, electrophoretic methods, RNase-digestion experiments, and its in vitro gene-silencing activity. The chemical synthesis of RNA oligomers of more than 100 nucleotides, which has until now been extremely difficult, can be practically realized by the CEM method.

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