posted on 2016-02-24, 00:00authored byBin Li, Xiao Luo, Yizhou Dong
Chemically
modified nucleotides play significant roles in the effectiveness
of mRNA translation. Here, we describe the synthesis of two sets of
chemically modified mRNAs [encoding firefly Luciferase (FLuc) and
enhanced green fluorescent protein (eGFP), respectively], evaluation
of protein expression, and correlation analysis of expression level
under various conditions. The results indicate that chemical modifications
of mRNAs are able to significantly improve protein expression, which
is dependent on cell types and coding sequences. Moreover, eGFP mRNAs
with N1-methylpseudouridine (me1ψ), 5-methoxyuridine (5moU), and pseudouridine (ψ) modifications
ranked top three in cell lines tested. Interestingly, 5moU-modified
eGFP mRNA was more stable than other eGFP mRNAs. Consequently, me1ψ, 5moU, and ψ are promising nucleotides for chemical
modification of mRNAs.