Most eukaryotic mRNAs require a 7-methyl guanosine (m7G) cap structure atthe 5´ end and a Poly(A) tail at the 3´ end to be efficiently translated. By usinga DNA template encoding a poly(A) tail, the HiScribe T7 ARCA mRNA Kit canbe used to synthesize capped and tailed mRNAs. The cap structure is addedto the mRNA by co-transcriptional incorporation of Anti-Reverse Cap Analog(ARCA) (NEB #S1411) using T7 RNA Polymerase. Poly(A) tail is incorporatedduring the transcription reaction. The kit also includes DNase I and LiCl forDNA template removal and quick mRNA purification.Additionally, the kit is capable of partial incorporation of modified UTP andCTP (up to 50% each) without affecting the mRNA yield significantly. By usinga DNA template encoding a poly(A) tail, capped and tailed modified mRNA canbe synthesized in a single reaction in 30 minutes. mRNAs synthesized withthe kit can be used for cell transfection, microinjection, in vitro translation andRNA vaccines.ARCA is incorporated into mRNA exclusively in the correct orientation, generatingcapped mRNA that is more efficiently translated. Standard cap analogscan be incorporated in either direction resulting in only 50% of capped mRNAthat is functional in protein translation.Figure 1. Structure of Anti-Reverse Cap Analog (ARCA, NEB #S1411)Methylation at the 3´ position of 7mG forces the cap structure to be attached to mRNA in the correct orientation.Figure 2. Overview of mRNA synthesis work flow with the HiScribeT7 ARCA mRNA Kit
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RNA Synthesis Products
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