Title:
ENZYMATIC RNA SYNTHESIS
Document Type and Number:
WIPO Patent Application WO/2020/077227
Kind Code:
A3
Abstract:
Described herein are methods for the controlled de novo synthesis of RNA oligonucleotides using enzymatic catalysis. For example, provided herein are methods for preparing RNA oligonucleotides via controlled, template-independent addition of nucleotides to an initiator oligonucleotide 3'- terminus via enzymatic catalysis (also known as terminal transferase activity). Single nucleotides can be iteratively added by a compatible polymerase (e.g., a poly(N) polymerase such as a poly(U) polymerase) until a desired RNA oligonucleotide sequence is synthesized. Also provided are nucleotides and polymerases useful in the methods described herein.
Inventors:
CHURCH GEORGE (US)
WIEGAND DANIEL (US)
KOHMAN RICHARD (US)
KURU ERKIN (US)
RITTICHIER JONATHAN (US)
CONWAY NICHOLAS (US)
WIEGAND DANIEL (US)
KOHMAN RICHARD (US)
KURU ERKIN (US)
RITTICHIER JONATHAN (US)
CONWAY NICHOLAS (US)
Application Number:
PCT/US2019/055870
Publication Date:
August 27, 2020
Filing Date:
October 11, 2019
Export Citation:
Assignee:
HARVARD COLLEGE (US)
International Classes:
C12N15/11; C12N9/12; C12P19/34
Domestic Patent References:
| WO2005118806A2 | 2005-12-15 | |||
| WO2011038241A1 | 2011-03-31 |
Foreign References:
| EP2551354A1 | 2013-01-30 | |||
| US20160046974A1 | 2016-02-18 |
Other References:
MARIE-LUISE WINZ ET AL: "Site-specific terminal and internal labeling of RNA by poly(A) polymerase tailing and copper-catalyzed or copper-free strain-promoted click chemistry", NUCLEIC ACIDS RESEARCH, vol. 40, no. 10, 16 February 2012 (2012-02-16), GB, pages e78 - e78, XP055441625, ISSN: 0305-1048, DOI: 10.1093/nar/gks062
DANIEL DEHANY SCOTT: "Characterisation of RNA uridylyltransferases in Schizosaccharomyces pombe", PH. D. THESIS, 2012, XP055677469, Retrieved from the Internet [retrieved on 20200318]
PAOLA MUNOZ-TELLO ET AL: "Functional Implications from the Cid1 Poly(U) Polymerase Crystal Structure", STRUCTURE, ELSEVIER, AMSTERDAM, NL, vol. 20, no. 6, 17 May 2012 (2012-05-17), pages 977 - 986, XP028520133, ISSN: 0969-2126, [retrieved on 20120426], DOI: 10.1016/J.STR.2012.04.006
MATULIC-ADAMIC JASENKA ET AL: "Functionalized nucleoside 5'-triphosphates for in vitro selection of new catalytic ribonucleic acids", BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, PERGAMON, AMSTERDAM, NL, vol. 10, no. 11, 9 June 2000 (2000-06-09), pages 1299 - 1302, XP085058276, ISSN: 0960-894X, DOI: 10.1016/S0960-894X(00)00226-2
B. MORITZ ET AL: "Simple methods for the 3' biotinylation of RNA", RNA, vol. 20, no. 3, 21 January 2014 (2014-01-21), US, pages 421 - 427, XP055354650, ISSN: 1355-8382, DOI: 10.1261/rna.042986.113
YUNLEI ZHOU ET AL: "Electrochemical biosensor for microRNA detection based on poly(U) polymerase mediated isothermal signal amplification", BIOSENSORS AND BIOELECTRONICS, vol. 79, 9 December 2015 (2015-12-09), AMSTERDAM, NL, pages 79 - 85, XP055677399, ISSN: 0956-5663, DOI: 10.1016/j.bios.2015.12.009
DANIEL DEHANY SCOTT: "Characterisation of RNA uridylyltransferases in Schizosaccharomyces pombe", PH. D. THESIS, 2012, XP055677469, Retrieved from the Internet
PAOLA MUNOZ-TELLO ET AL: "Functional Implications from the Cid1 Poly(U) Polymerase Crystal Structure", STRUCTURE, ELSEVIER, AMSTERDAM, NL, vol. 20, no. 6, 17 May 2012 (2012-05-17), pages 977 - 986, XP028520133, ISSN: 0969-2126, [retrieved on 20120426], DOI: 10.1016/J.STR.2012.04.006
MATULIC-ADAMIC JASENKA ET AL: "Functionalized nucleoside 5'-triphosphates for in vitro selection of new catalytic ribonucleic acids", BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, PERGAMON, AMSTERDAM, NL, vol. 10, no. 11, 9 June 2000 (2000-06-09), pages 1299 - 1302, XP085058276, ISSN: 0960-894X, DOI: 10.1016/S0960-894X(00)00226-2
B. MORITZ ET AL: "Simple methods for the 3' biotinylation of RNA", RNA, vol. 20, no. 3, 21 January 2014 (2014-01-21), US, pages 421 - 427, XP055354650, ISSN: 1355-8382, DOI: 10.1261/rna.042986.113
YUNLEI ZHOU ET AL: "Electrochemical biosensor for microRNA detection based on poly(U) polymerase mediated isothermal signal amplification", BIOSENSORS AND BIOELECTRONICS, vol. 79, 9 December 2015 (2015-12-09), AMSTERDAM, NL, pages 79 - 85, XP055677399, ISSN: 0956-5663, DOI: 10.1016/j.bios.2015.12.009
Attorney, Agent or Firm:
HAMZIK, Philip, J. et al. (US)
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