Solid phase synthesis of mRNA by the phosphoramidite approach using 2'-O-1-(2-chloroethoxy)ethyl protection and its stability in E. coli system.
The decaoligoribonucleotides containing initiation codon AUG of the phage O beta-A protein mRNA were synthesized on a solid phase by the phosphoramidite approach using 2'-O-1-(2-chloroethoxy)ethyl (Cee) protection. The Cee group is completely stable under the acidic conditions required to remove the 5'-terminal protecting groups in oligoribonucleotide synthesis on a solid support, and yet is easily removable at pH 2.0 for the final unblocking step. Stabilization of the synthetic mRNA to in the cell-free translation system from E. coli A19 was measured. It was found that the oligomers with selected 2'-O-methylation were degraded completely in the translation system.
Sakatsume
O
O
Department of Industrial Chemistry, Chiba Institute of Technology, Japan.
Yamaguchi
T
T
Takaku
H
H
D001483:Base Sequence; D003062:Codon; D004926:Escherichia coli / Q000378:metabolism*; D004987:Ethers; D006868:Hydrolysis; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D009843:Oligoribonucleotides / Q000138:chemical synthesis*; D009943:Organophosphorus Compounds; D012333:RNA, Messenger / Q000138:chemical synthesis*
D003062:Codon; D004987:Ethers; D007202:Indicators and Reagents; D009843:Oligoribonucleotides; D009943:Organophosphorus Compounds; D012333:RNA, Messenger