Chemical synthesis of a very long RNA oligomer, a 110mer precursor-miRNA candidate, with 2-cyanoethoxymethyl (CEM) as the 2'-O-protecting group.
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.
Masuda
Hirofumi
H
Discovery Research Laboratories, Nippon Shinyaku Co. Ltd., 3-14-1 Sakura, Tsukuba City, Ibaraki 305-0003, Japan.
Shiba
Yoshinobu
Y
Watanabe
Naoki
N
Takagaki
Kazuchika
K
Ishiyama
Kouichi
K
Ohgi
Tadaaki
T
Yano
Junichi
J
D001671:Biochemistry / Q000379:methods; D005019:Ethyl Ethers / Q000737:chemistry*; D008738:Methyl Ethers / Q000737:chemistry*; D035683:MicroRNAs / Q000138:chemical synthesis* / Q000737:chemistry; D009843:Oligoribonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D034622:RNA Interference; D012322:RNA Precursors / Q000138:chemical synthesis* / Q000737:chemistry
C522071:2-cyanoethyl chloromethyl ether; D005019:Ethyl Ethers; D008738:Methyl Ethers; D035683:MicroRNAs; D009843:Oligoribonucleotides; D012322:RNA Precursors