Title:
CONTROLLED TEMPLATE-INDEPENDENT SYNTHESIS OF NUCLEIC ACIDS USING THERMOSTABLE ENZYMES
Document Type and Number:
WIPO Patent Application WO/2021/250269
Kind Code:
A3
Abstract:
The invention relates to methods for template-independent synthesis of nucleic acids, comprising iteratively contacting an initiator sequence comprising a 3'-end nucleotide with a free 3'-hydroxyl group, with at least one nucleoside triphosphate, or a combination of nucleoside triphosphates, in the presence of an archaeal DNA primase or a functionally active fragment and/or variant thereof, thereby covalently binding said nucleoside triphosphate to the free 3'-hydroxyl group of the 3'-end nucleotide. It also relates to isolated functionally active fragments of archaeal DNA primases which are capable of template-independent terminal nucleotidyl transferase activity but are devoid of a template-independent primase activity.
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Inventors:
RANDRIANJATOVO-GBALOU IRINA (FR)
SAID AHMED (FR)
RAHIER RENAUD (FR)
SAID AHMED (FR)
RAHIER RENAUD (FR)
Application Number:
PCT/EP2021/065867
Publication Date:
February 10, 2022
Filing Date:
June 11, 2021
Export Citation:
Assignee:
SYNHELIX (FR)
International Classes:
C12N9/12; C12P19/34; C12Q1/6806
Foreign References:
| US20190323050A1 | 2019-10-24 | |||
| US20190360013A1 | 2019-11-28 | |||
| US20190078126A1 | 2019-03-14 | |||
| EP2357225A1 | 2011-08-17 |
Other References:
SUKHVINDER GILL ET AL: "A highly divergent archaeo-eukaryotic primase from the Thermococcus nautilus plasmid, pTN2", NUCLEIC ACIDS RESEARCH, vol. 42, no. 6, 20 January 2014 (2014-01-20), GB, pages 3707 - 3719, XP055605680, ISSN: 0305-1048, DOI: 10.1093/nar/gkt1385
BÉGUIN PIERRE ET AL: "Synergistic template-free synthesis of dsDNA byThermococcus nautiliprimase PolpTN2, DNA polymerase PolB, and pTN2 helicase", EXTREMOPHILES, SPRINGER VERLAG, TOKYO, JP, vol. 19, no. 1, 25 November 2014 (2014-11-25), pages 69 - 76, XP055764476, ISSN: 1431-0651, [retrieved on 20141125], DOI: 10.1007/S00792-014-0706-1
KRUPOVIC MART ET AL: "Insights into Dynamics of Mobile Genetic Elements in Hyperthermophilic Environments from Five New Thermococcus Plasmids", vol. 8, no. 1, 11 January 2013 (2013-01-11), pages e49044, XP055836613, Retrieved from the Internet DOI: 10.1371/journal.pone.0049044
BÉGUIN PIERRE ET AL: "Synergistic template-free synthesis of dsDNA byThermococcus nautiliprimase PolpTN2, DNA polymerase PolB, and pTN2 helicase", EXTREMOPHILES, SPRINGER VERLAG, TOKYO, JP, vol. 19, no. 1, 25 November 2014 (2014-11-25), pages 69 - 76, XP055764476, ISSN: 1431-0651, [retrieved on 20141125], DOI: 10.1007/S00792-014-0706-1
KRUPOVIC MART ET AL: "Insights into Dynamics of Mobile Genetic Elements in Hyperthermophilic Environments from Five New Thermococcus Plasmids", vol. 8, no. 1, 11 January 2013 (2013-01-11), pages e49044, XP055836613, Retrieved from the Internet
Attorney, Agent or Firm:
ICOSA (FR)
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