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

• Published in: Bulletin of experimental biology and medicine Vol. 176; no. 6; pp. 751 - 755
• Main Authors: Sharabrin, S. V., Bondar, A. A., Starostina, E. V., Kisakov, D. N., Kisakova, L. A., Zadorozhny, A. M., Rudometov, A. P., Ilyichev, A. A., Karpenko, L. I.
• Format: Journal Article
• Language: English
• Published: New York Springer US 01.04.2024; Springer Nature B.V
• Subjects: Antibodies; Biology; Biomedical and Life Sciences; Biomedicine; Biotechnology; CD69 antigen; Cell activation; Cell Biology; Cellulose; Contaminants; DNA-directed RNA polymerase; Double-stranded RNA; Impurities; Inflammation; Internal Medicine; Kinases; Laboratory Medicine; Lymphocytes; mRNA vaccines; Pathology; Proteins; Research centers; RNA polymerase; Toxicology; Transcription; Virology; α-Interferon; dsRNA; mRNA purification; mRNA
• ISSN: 0007-4888; 1573-8221; 1573-8221
• DOI: 10.1007/s10517-024-06102-2

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.