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

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

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.

Afiliación

Sharabrin SV; State Research Center of Virology and Biotechnology "VECTOR", Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, Koltsovo, Novosibirsk region, Russia. sharabrin.sv@gmail.com.
Bondar AA; Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia.
Starostina EV; State Research Center of Virology and Biotechnology "VECTOR", Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, Koltsovo, Novosibirsk region, Russia.
Kisakov DN; State Research Center of Virology and Biotechnology "VECTOR", Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, Koltsovo, Novosibirsk region, Russia.
Kisakova LA; State Research Center of Virology and Biotechnology "VECTOR", Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, Koltsovo, Novosibirsk region, Russia.
Zadorozhny AM; State Research Center of Virology and Biotechnology "VECTOR", Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, Koltsovo, Novosibirsk region, Russia.
Rudometov AP; State Research Center of Virology and Biotechnology "VECTOR", Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, Koltsovo, Novosibirsk region, Russia.
Ilyichev AA; State Research Center of Virology and Biotechnology "VECTOR", Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, Koltsovo, Novosibirsk region, Russia.
Karpenko LI; State Research Center of Virology and Biotechnology "VECTOR", Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing, Koltsovo, Novosibirsk region, Russia.

Bull Exp Biol Med ; 176(6): 751-755, 2024 Apr.

Article en En | MEDLINE | ID: mdl-38896322

ABSTRACT

ABSTRACT

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.

Asunto(s)

ARN Bicatenario; ARN Mensajero; Animales; ARN Mensajero/genética; ARN Mensajero/metabolismo; ARN Bicatenario/genética; ARN Bicatenario/metabolismo; Ratones; Antígenos de Diferenciación de Linfocitos T/metabolismo; Antígenos de Diferenciación de Linfocitos T/genética; Antígenos CD/genética; Antígenos CD/metabolismo; Lectinas Tipo C/metabolismo; Lectinas Tipo C/genética; ARN Polimerasas Dirigidas por ADN/metabolismo; ARN Polimerasas Dirigidas por ADN/genética; Interferón-alfa/genética; Interferón-alfa/metabolismo; Interferón-alfa/biosíntesis; Proteínas Virales/metabolismo; Proteínas Virales/genética

Palabras clave

dsRNA; mRNA; mRNA purification

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Texto completo: 1 Colección: 01-internacional Banco de datos: MEDLINE Asunto principal: ARN Bicatenario / ARN Mensajero Límite: Animals Idioma: En Revista: Bull Exp Biol Med Año: 2024 Tipo del documento: Article País de afiliación: Rusia

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