The 5´terminal m7G cap present on most eukaryotic mRNAs promotes translation in vitro at the initiation level (1,2,3).
A method for efficient in vitro synthesis of capped RNA using E.coli RNA polymerase primed with m7G(5´ )ppp(5´ )G (#S1404S) or m7G(5´ )ppp(5´ )A (#S1405S) has been developed by Contreas et al. (7).
For most RNAs, elimination of the cap structure causes a loss of stability, especially against exonuclease degradation (4), and a decrease in the formation of the initiation complex of mRNAs for protein synthesis (4,5). Certain prokaryotic mRNAs containing a 5´ terminal cap structure are translated as efficiently as or more efficiently than eukaryotic mRNAs in a eukaryotic cell-free protein synthesizing system (5). Also a cap requirement has been observed for splicing eukaryotic substrate RNAs (6).
Figure 1: Structure of m7G(5´)ppp(5´)G RNA Cap Structure Analog (NEB #S1404)
Sequence
7mG(5´)ppp(5´)G
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