T7 ARCA加帽 mRNA合成試劑盒
It produces capped RNA by in vitro transcription using T7 RNA polymerase and the anti-reverse cap analog (ARCA) m2 7,3'-OG[5']ppp[5'].
Product Name：T7 ARCA-Capped Message Transcription Kit(T7 ARCA加帽 mRNA合成試劑盒)
Introduction：
The MessageMAX™ T7 ARCA-Capped Message Transcription Kit produces capped RNA by in vitro transcription using T7 RNA polymerase and the anti-reverse cap analog (ARCA) m2 7,3'-OG[5']ppp[5']. A 30 minute, 20 μl MessageMAX reaction yields up to 60 μg of ARCA-capped RNA from 1 μg of the 1.4 kb standard T7 control template DNA. Because ARCA contains a 3'-O-methyl group on the m7 G nucleotide (Figure 1), ARCA can only be incorporated in the correct orientation at the 5' end of the RNA during an in vitro transcription/capping reaction.1-3 This is not true for the standard cap analog (m7 G[5']ppp[5']G). Thus, ARCA incorporation results in the synthesis of capped RNA that is more efficiently translated in vivo than standard cap analog. The ARCA/NTP PreMix contains all four ribonucleotides and the ARCA. The PreMix ensures the optimal concentration of each NTP and ratio of ARCA to GTP (4:1), maximizing transcript capping (~80%) and yield. Because the concentration of GTP in the reaction is limiting, the ARCA is preferentially incorporated as the first or 5'-terminal G of the transcript. CELLSCRIPT also offers the INCOGNITO™ T7 ARCA 5mC- & -RNA Transcription Kit for the production of 5-methylcytosine- pseudouridine-containing (GA m5 C) ARCA-capped RNA. Such RNA has been used in the landmark publication by Warren et al. (2010), to successfully reprogram multiple human cell types into induced pluripotent stem cells.4 It has been shown that -mRNAs and 5mCmRNAs are translated into protein at higher levels and induce lower innate immune responses in human and other mammalian cells that express various RNA sensors compared to corresponding unmodified mRNAs.
