Title:
METHOD FOR DETECTING PROPORTION OF MRNA WITH POLYA TAIL IN MRNA PRODUCT
Document Type and Number:
WIPO Patent Application WO/2024/051832
Kind Code:
A1
Abstract:
Disclosed in the present invention is a method for detecting the proportion of mRNA with a polyA tail in a mRNA product. The method comprises the following steps: 1) obtaining an RNA with an adapter at the 3' end; 2) performing reverse transcription to obtain a cDNA; 3) designing and synthesizing a primer and a probe for the 5' end of the cDNA, designing and synthesizing a primer and a probe for the 3' end of the cDNA, preparing a digital PCR reaction system for determination, and calculating the proportion of the mRNA with the polyA tail according to the copy number determined by the digital PCR.
Inventors:
HU YONG (CN)
TANG ZAN (CN)
WANG RUIYUE (CN)
XU LIANQIANG (CN)
WANG SHUAI (CN)
TANG ZAN (CN)
WANG RUIYUE (CN)
XU LIANQIANG (CN)
WANG SHUAI (CN)
Application Number:
PCT/CN2023/117779
Publication Date:
March 14, 2024
Filing Date:
September 08, 2023
Export Citation:
Assignee:
SHENZHEN RHEGEN BIOTECHNOLOGY CO LTD (CN)
WUHAN RHEGEN BIOTECHNOLOGY CO LTD (CN)
WUHAN RHEGEN BIOTECHNOLOGY CO LTD (CN)
International Classes:
C12Q1/68; C12Q1/6851
Foreign References:
| CN102277351A | 2011-12-14 | |||
| US20200325532A1 | 2020-10-15 | |||
| US20090069194A1 | 2009-03-12 |
Other References:
BROUZE ALEKSANDRA, KRAWCZYK PAWEĊ SZCZEPAN, DZIEMBOWSKI ANDRZEJ, MROCZEK SEWERYN: "Measuring the tail: Methods for poly(A) tail profiling", WILEY INTERDISCIPLINARY REVIEWS: RNA, JOHN WILEY & SONS LTD, UNITED KINGDOM, vol. 14, no. 1, 1 January 2023 (2023-01-01), United Kingdom , pages e1737, XP093146884, ISSN: 1757-7004, DOI: 10.1002/wrna.1737
MICHAEL BEVERLY ET AL.: "Poly A tail length analysis of in vitro transcribed mRNA by LC-MS", ANALYTICAL AND BIOANALYTICAL CHEMISTRY, vol. 410, no. 6, 8 January 2018 (2018-01-08), XP036416023, DOI: 10.1007/s00216-017-0840-6
ELIZABETH L. MURRAY, SCHOENBERG DANIEL R. : "Assays for determining poly(A) tail length and the polarity of mRNA decay in mammalian cells ", METHODS ENZYMOL., vol. 448, 1 January 2008 (2008-01-01), pages 483 - 504, XP055049447, DOI: 10.1016/S0076-6879(08)02624-4
MICHAEL BEVERLY ET AL.: "Poly A tail length analysis of in vitro transcribed mRNA by LC-MS", ANALYTICAL AND BIOANALYTICAL CHEMISTRY, vol. 410, no. 6, 8 January 2018 (2018-01-08), XP036416023, DOI: 10.1007/s00216-017-0840-6
ELIZABETH L. MURRAY, SCHOENBERG DANIEL R. : "Assays for determining poly(A) tail length and the polarity of mRNA decay in mammalian cells ", METHODS ENZYMOL., vol. 448, 1 January 2008 (2008-01-01), pages 483 - 504, XP055049447, DOI: 10.1016/S0076-6879(08)02624-4
Attorney, Agent or Firm:
BEIJING SANYOU INTELLECTUAL PROPERTY AGENCY LTD. (CN)
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