Q: What would be the advantage of using modified nucleotides?
A: mRNAs modified with the incorporation of N1meΨTP and ΨTP have demonstrated the ability to reduce RNA-mediated innate immune activation in vivo.
Q: Are modified nucleotides included in the INCOGNITO™ T7 mScript™ mRNA Production Systems?
A: Yes, the modified nucleotides N1meΨTP and ΨTP are included in their respective kits.
Q: Which process has better capping efficiency, the INCOGNITO™ T7 mScript™ mRNA Production Systems from CELLSCRIPT™ or traditional G-start cap analog-based co-transcriptional capping systems such as your MessageMAX™ T7 ARCA-Capped Message Transcription Kit V2?
A: The INCOGNITO™ T7 mScript™ mRNA Production Systems produce capped mRNA with nearly 100% efficiency and with all the caps in the correct orientation. Direct incorporation of cap analog dinucleotides during traditional G-start co-transcriptional systems is considerably less efficient, where capping efficiencies cannot exceed 80%.
Q: Why is there such a large improvement in RNA capping efficiency using the INCOGNITO™ T7 mScript™ mRNA Production Systems when compared to traditional G-start cap analog-based co-transcriptional capping reactions?
A: Unlike traditional G-start cap analog-based co-transcriptional reactions the INCOGNITO™ T7 mScript™ mRNA Production Systems use an enzymatic process to build cap structures onto the RNA transcripts which bypasses several of the limitations of the traditional G-start cap analog-based chemistry. These limitations include lower capping efficiencies, reduced in vitro transcription (IVT) reaction yields and limited IVT RNA lengths. The INCOGNITO™ T7 mScript™ mRNA Production Systems include ScriptCap™ Capping Enzyme, which contains all three enzymatic activities (mRNA triphosphatase, guanylyltransferase and guanine-7-methyltransferase) necessary to build a 5′-Cap 0 structure in vitro onto the RNA, ensuring nearly 100% capping with 100% proper cap orientation. The INCOGNITO™ T7 mScript™ mRNA Production Systems also contain ScriptCap™ 2′-O-Methyltransferase, allowing the natural, translation boosting and self RNA identifying, Cap 1 structure to be built onto the RNA.
