Title:
METHOD FOR PRODUCING NUCLEIC ACID MOLECULE
Document Type and Number:
WIPO Patent Application WO/2021/024465
Kind Code:
A1
Abstract:
The present invention provides a method for producing single-stranded RNA, comprising the steps of: (I) allowing a RNA ligase that is classified into an enzyme commission number of EC6.5.1.3 as assigned by the International Union of Biochemistry and has an activity to repair double-strand nicks to act on first single-stranded RNA that has a phosphate group at the 5'-terminal thereof and second single-stranded RNA that has a hydroxyl group at the 3'-terminal thereof to connect the first single-stranded RNA to the second single-stranded RNA, thereby producing single-stranded RNA; and (II) purifying a reaction product that contains the single-stranded RNA produced in the connection step (I) by reverse-phase column chromatography using a mobile phase containing at least one ammonium salt selected from the group consisting of a monoalkyl ammonium salt and a dialkyl ammonium salt.
Inventors:
SAKATA AKIHIRO (JP)
Application Number:
PCT/JP2019/031421
Publication Date:
February 11, 2021
Filing Date:
August 08, 2019
Export Citation:
Assignee:
SUMITOMO CHEMICAL CO (JP)
International Classes:
C12N15/10; C12N15/11; C12P19/34
Domestic Patent References:
| WO2019156020A1 | 2019-08-15 |
Foreign References:
| JP2017046596A | 2017-03-09 | |||
| US20170121716A1 | 2017-05-04 |
Other References:
YIN S. ET AL.: "Characterization of bacteriophage KVP40 and T4 RNA ligase 2", VIROLOGY, vol. 319, no. 1, 2004, pages 141 - 151, XP004488998, DOI: 10.1016/j.virol.2003.10.037
MARTINS A ET AL.: "An RNA Ligase from Deinococcus radiodurans", THE JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 279, no. 49, 2004, pages 50654 - 50661, XP055629625, DOI: 10.1074/jbc.M407657200
GONG, L.: "Comparing ion-pairing reagents and counter anions for ion-pair reversed-phase liquid chromatography/electrospray ionization mass spectrometry analysis of synthetic oligonucleotides", RAPID COMMUNICATIONS IN MASS SPECTROMETRY, vol. 29, 2015, pages 2402 - 2410, XP055354828, DOI: 10.1002/rcm.7409
GONG, L. ET AL.: "Comparing ion-pairing reagents and sample dissolution solvents for ion-pairing reversed-phase liquid chromatography/electrospray ionization mass spectrometry analysis of oligonucleotides", RAPID COMMUNICATIONS IN MASS SPECTROMETRY, vol. 28, 2014, pages 339 - 350, XP055354822, DOI: 10.1002/rcm.6773
MARTINS A ET AL.: "An RNA Ligase from Deinococcus radiodurans", THE JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 279, no. 49, 2004, pages 50654 - 50661, XP055629625, DOI: 10.1074/jbc.M407657200
GONG, L.: "Comparing ion-pairing reagents and counter anions for ion-pair reversed-phase liquid chromatography/electrospray ionization mass spectrometry analysis of synthetic oligonucleotides", RAPID COMMUNICATIONS IN MASS SPECTROMETRY, vol. 29, 2015, pages 2402 - 2410, XP055354828, DOI: 10.1002/rcm.7409
GONG, L. ET AL.: "Comparing ion-pairing reagents and sample dissolution solvents for ion-pairing reversed-phase liquid chromatography/electrospray ionization mass spectrometry analysis of oligonucleotides", RAPID COMMUNICATIONS IN MASS SPECTROMETRY, vol. 28, 2014, pages 339 - 350, XP055354822, DOI: 10.1002/rcm.6773
Attorney, Agent or Firm:
YAMAO, Norihito et al. (JP)
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