Title:
METHOD FOR CELL-FREE PREPARATION OF MRNA TRANSCRIPTION TEMPLATE
Document Type and Number:
WIPO Patent Application WO/2024/067644
Kind Code:
A1
Abstract:
A method for cell-free preparation of an mRNA transcription template, the method comprising the following steps: 1) in the presence of at least one type of primer and while promoting the amplification of a DNA template, causing contact of a circular DNA template comprising at least one palindromic sequence with at least one type of DNA polymerase having strand displacement activity; and 2) causing contact of the DNA product prepared in step 1 with at least one type of restriction enzyme. Compared to PCR amplification methods, the present method for cell-free preparation of an mRNA transcription template can achieve large-scale preparation; compared to microbial fermentation methods,the present method had added polyA stability and does not involve culturing of live organisms, which simplifies the preparation process, and is a superior method for preparing a DNA template in the field of mRNA.
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Inventors:
HU YONG (CN)
ZHANG XING (CN)
AI LIANGXIA (CN)
XIAO XUETING (CN)
XU PAN (CN)
LYU BIN (CN)
ZHANG XING (CN)
AI LIANGXIA (CN)
XIAO XUETING (CN)
XU PAN (CN)
LYU BIN (CN)
Application Number:
PCT/CN2023/121770
Publication Date:
April 04, 2024
Filing Date:
September 26, 2023
Export Citation:
Assignee:
SHENZHEN RHEGEN BIOTECHNOLOGY CO LTD (CN)
WUHAN RHEGEN BIOTECHNOLOGY CO LTD (CN)
WUHAN RHEGEN BIOTECHNOLOGY CO LTD (CN)
International Classes:
C12P19/34; C12N9/16; C12N9/22; C12N15/10; C12Q1/68; C12Q1/6844
Foreign References:
| CN103080337A | 2013-05-01 | |||
| CN104232628A | 2014-12-24 | |||
| CN101802183A | 2010-08-11 | |||
| CN110484606A | 2019-11-22 |
Other References:
HOLTKAMP, S. ET AL.: "Modification of antigen-encoding RNA increases stability, translational efficacy, and T-cell stimulatory capacity of dendritic cells", BLOOD., vol. 108, no. 13, 15 December 2006 (2006-12-15), XP055044965, DOI: 10.1182/blood-2006-04-015024
JOFFROY, B. ET AL.: "Rolling circle amplification shows a sinusoidal template length-dependent amplification bias", NUCLEIC ACIDS RES., vol. 46, no. 2, 25 January 2018 (2018-01-25), XP055659491, DOI: 10.1093/nar/gkx1238
JOFFROY, B. ET AL.: "Rolling circle amplification shows a sinusoidal template length-dependent amplification bias", NUCLEIC ACIDS RES., vol. 46, no. 2, 25 January 2018 (2018-01-25), XP055659491, DOI: 10.1093/nar/gkx1238
Attorney, Agent or Firm:
BEIJING SANYOU INTELLECTUAL PROPERTY AGENCY LTD. (CN)
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