ABSTRACT
Little is known about potential protective factors for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), referred to as COVID-19. Suboptimal vitamin D status is a risk factor for immune dysfunction, respiratory tract infections (RTIs), and viral infections. Supplementation of vitamin D (2000-4000 IU) has decreased incidence and complications from RTIs, respiratory distress syndrome, and pneumonia and may be beneficial in high-risk populations. Given the possible link between low vitamin D status and RTIs, such as COVID-19, this review examined whether vitamin D supplementation can be supported as a nutritional strategy for reducing risk of infection, complications, and mortality from COVID-19 and found that the relationship between vitamin D and RTIs warrants further exploration.
ABSTRACT
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. © 2021 Wiley Periodicals LLC. Basic Protocol 1: IVT with CleanCap Basic Protocol 2: mRNA purification and analysis.