Title:
METHOD FOR PREPARING RECOMBINANT EUKARYOTIC MRNA BY USING PROKARYOTIC TRANSCRIPTION SYSTEM AND APPLICATION THEREOF
Document Type and Number:
WIPO Patent Application WO/2021/259057
Kind Code:
A1
Abstract:
Provided is a method for preparing a recombinant eukaryotic mRNA molecule by using a prokaryotic transcription system. The method comprises: 1) expressing an enzyme system required for the transcription and capping process of the recombinant eukaryotic mRNA, and obtaining a prokaryotic in-vitro transcription-capping enzyme system; 2) preparing a linearized transcription template DNA; and 3) transcribing, capping and purifying the recombinant mRNA. The prepared recombinant mRNA molecule has all mRNA characteristics required by eukaryotic organisms in a protein translation process, and therefore, the recombinant mRNA molecule can be further identified and utilized by a eukaryotic translation system.
Inventors:
CHEN BO (CN)
LI RAN (CN)
WANG RUKAI (CN)
LI RAN (CN)
WANG RUKAI (CN)
Application Number:
PCT/CN2021/098748
Publication Date:
December 30, 2021
Filing Date:
June 07, 2021
Export Citation:
Assignee:
WUHAN DANGKANG XING ZHONG BIOTECHNOLOGY CO LTD (CN)
International Classes:
C12N15/10; C12N15/52; C12N15/54; C12N15/63; C12N15/70
Domestic Patent References:
| WO2014028429A2 | 2014-02-20 |
Foreign References:
| CN109104870A | 2018-12-28 | |||
| CN111893128A | 2020-11-06 | |||
| US20190062762A1 | 2019-02-28 |
Other References:
FUCHS ANNA-LISA, NEU ANCILLA, SPRANGERS REMCO: "A general method for rapid and cost-efficient large-scale production of 5′ capped RNA", RNA, COLD SPRING HARBOR LABORATORY PRESS, US, vol. 22, no. 9, 1 September 2016 (2016-09-01), US , pages 1454 - 1466, XP055882698, ISSN: 1355-8382, DOI: 10.1261/rna.056614.116
SHUMAN STEWART: "Catalytic activity of vaccinia mRNA capping enzyme subunits coexpressed in Escherichia coli.", THE JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 265, no. 20, 15 July 1990 (1990-07-15), pages 11960 - 11966, XP055882701, DOI: 10.1016/S0021-9258(19)38494-7
ANAND RAMANATHAN, G. BRETT ROBB, SIU-HONG CHAN: "mRNA capping: biological functions and applications", NUCLEIC ACIDS RESEARCH, OXFORD UNIVERSITY PRESS, GB, vol. 44, no. 16, 19 September 2016 (2016-09-19), GB , pages 7511 - 7526, XP055388045, ISSN: 0305-1048, DOI: 10.1093/nar/gkw551
HU SHUN, YI YOU-JIN;HU TAO;LI FU-SHENG: "Development and Clinical Progress of mRNA Vaccine", CHINA BIOTECHNOLOGY, vol. 39, no. 11, 23 April 2019 (2019-04-23), pages 105 - 112, XP055882709, DOI: 10.13523/j.cb.20191112
SHUMAN STEWART: "Catalytic activity of vaccinia mRNA capping enzyme subunits coexpressed in Escherichia coli.", THE JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 265, no. 20, 15 July 1990 (1990-07-15), pages 11960 - 11966, XP055882701, DOI: 10.1016/S0021-9258(19)38494-7
ANAND RAMANATHAN, G. BRETT ROBB, SIU-HONG CHAN: "mRNA capping: biological functions and applications", NUCLEIC ACIDS RESEARCH, OXFORD UNIVERSITY PRESS, GB, vol. 44, no. 16, 19 September 2016 (2016-09-19), GB , pages 7511 - 7526, XP055388045, ISSN: 0305-1048, DOI: 10.1093/nar/gkw551
HU SHUN, YI YOU-JIN;HU TAO;LI FU-SHENG: "Development and Clinical Progress of mRNA Vaccine", CHINA BIOTECHNOLOGY, vol. 39, no. 11, 23 April 2019 (2019-04-23), pages 105 - 112, XP055882709, DOI: 10.13523/j.cb.20191112
Attorney, Agent or Firm:
SUZHOU HUABO INTELLECTUAL PROPERTY AGENCY CO., LTD. (CN)
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