The 5´terminal m7G cap present on most eukaryotic mRNAs promotes translationin vitroat the initiation level (1,2,3). 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).
A method for efficientin vitrosynthesis of capped RNA usingE.coliRNA polymerase primed with m7G(5´ )ppp(5´ )G (#S1404S) or m7G(5´ )ppp(5´ )A (#S1405S) has been developed by Contreas et al. (7).