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首页> 外文期刊>Current Protocols in Nucleic Acid Chemistry >Cap 1 Messenger RNA Synthesis with Co-transcriptional CleanCap? Analog by In Vitro Transcription
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Cap 1 Messenger RNA Synthesis with Co-transcriptional CleanCap? Analog by In Vitro Transcription

机译：Cap 1 Messenger RNA Synthesis with Co-transcriptional CleanCap? Analog by In Vitro Transcription

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摘要

Synthetic messenger RNA (mRNA)-based therapeutics are an increasingly popular approach to gene and cell therapies, genome engineering, enzyme replacement therapy, and now, during the global SARS-CoV-2 pandemic, vaccine development. mRNA for such purposes can be synthesized through an enzymatic in vitro transcription (IVT) reaction and formulated for in vivo delivery. Mature mRNA requires a 5'-cap for gene expression and mRNA stability. There are two methods to add a cap in vitro: via a two-step multi-enzymatic reaction or co-transcriptionally. Co-transcriptional methods minimize reaction steps and enzymes needed to make mRNA when compared to enzymatic capping. CleanCap? AG co-transcriptional capping results in 5 mg/ml of IVT with 94% 5'-cap 1 structure. This is highly efficient compared to first-generation cap analogs, such as mCap and ARCA, that incorporate cap 0 structures at lower efficiency and reaction yield. This article describes co-transcriptional capping using TriLink Biotechnology's CleanCap? AG in IVT.

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