Removal of Double-Stranded RNA Contaminants During Template-Directed Synthesis of mRNA

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发表日期：

2024年4月1日

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S. V. Sharabrin,
A. A. Bondar,
E. V. Starostina,
D. N. Kisakov,
L. A. Kisakova,
A. M. Zadorozhny,
A. P. Rudometov,
A. A. Ilyichev,
L. I. Karpenko

摘要

The removal of double-stranded RNA (dsRNA) contaminants during in vitro mRNA synthesis is one of the technological problems to be solved. Apparently, these contaminants are the result of the T7 RNA polymerase side activity. In this study, we used a modified method of mRNA purification based on the selective binding of dsRNA to cellulose in ethanol-containing buffer. It was shown both in vivo and in vitro that the cellulose-purified mRNA preparation leads neither to activation of the lymphocyte inflammatory marker CD69 nor to increased release of IFNα in mice, and does not contain impurities detectable by antibodies to dsRNA.