The two faces of oligoadenylate synthetase-like: effective antiviral protein and negative regulator of innate immunity

Rex, Viktoria; Stempel, Markus; Halle, Stephan; Brinkmann, Melanie M.

doi:10.1016/j.coviro.2023.101329

Cited by 2 publications

(1 citation statement)

References 41 publications

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“…The minimum duplex lengths that are required for activation vary between ∼20bp for OAS1 to ∼35bp for OAS2 and ∼50bp for OAS3, even though the mechanism of dsRNA binding and activation is still unresolved ( Donovan et al, 2015 ; Koul et al, 2020 ; Wang et al, 2020 ). OASL, although also capable of binding dsRNA without having an enzymatic activity, is thought to influence the immune response through other pathways, like activation of RIG-I signaling, but mechanistic insights remain unclear ( Rex et al, 2023 ). Even though the binding mechanism is unknown, it was shown that RNA with modified nucleotides like pseudouridine, m6-adenosine and s2-uridine has a reduced capability to activate OASes and is resistant to degradation by RNase L ( Anderson et al, 2011 ).…”

Section: Ivt Byproducts and Their Biological Impactsmentioning

confidence: 99%

Understanding the impact of in vitro transcription byproducts and contaminants

Lenk,

Kleindienst,

Szabó

et al. 2024

Front. Mol. Biosci.

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The success of messenger (m)RNA-based vaccines against SARS-CoV-2 during the COVID-19 pandemic has led to rapid growth and innovation in the field of mRNA-based therapeutics. However, mRNA production, whether in small amounts for research or large-scale GMP-grade for biopharmaceutics, is still based on the In Vitro Transcription (IVT) reaction developed in the early 1980s. The IVT reaction exploits phage RNA polymerase to catalyze the formation of an engineered mRNA that depends on a linearized DNA template, nucleotide building blocks, as well as pH, temperature, and reaction time. But depending on the IVT conditions and subsequent purification steps, diverse byproducts such as dsRNA, abortive RNAs and RNA:DNA hybrids might form. Unwanted byproducts, if not removed, could be formulated together with the full-length mRNA and cause an immune response in cells by activating host pattern recognition receptors. In this review, we summarize the potential types of IVT byproducts, their known biological activity, and how they can impact the efficacy and safety of mRNA therapeutics. In addition, we briefly overview non-nucleotide-based contaminants such as RNases, endotoxin and metal ions that, when present in the IVT reaction, can also influence the activity of mRNA-based drugs. We further discuss current approaches aimed at adjusting the IVT reaction conditions or improving mRNA purification to achieve optimal performance for medical applications.

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