Title:
METHOD FOR PRODUCING CAS3 PROTEIN
Document Type and Number:
WIPO Patent Application WO/2022/186063
Kind Code:
A1
Abstract:
According to the present invention, it was discovered that a recombinant Cas3 protein can be efficiently expressed while maintaining the activity thereof by culturing, at a relatively low temperature, insect cells into which Cas3 gene is introduced, and that it is possible to recover an active form of the recombinant Cas3 protein with high purity at a high yield by purifying a solubilized fraction of the cells.
Inventors:
MASHIMO TOMOJI (JP)
YOSHIMI KAZUTO (JP)
TAKESHITA KOHEI (JP)
YAMAMOTO MASAKI (JP)
SHIBUMURA SATOMI (JP)
YOSHIMI KAZUTO (JP)
TAKESHITA KOHEI (JP)
YAMAMOTO MASAKI (JP)
SHIBUMURA SATOMI (JP)
Application Number:
PCT/JP2022/007821
Publication Date:
September 09, 2022
Filing Date:
February 25, 2022
Export Citation:
Assignee:
C4U CORP (JP)
UNIV TOKYO (JP)
RIKEN (JP)
UNIV TOKYO (JP)
RIKEN (JP)
International Classes:
C12P21/02; C07K14/195; C12N9/16; C12N15/31
Domestic Patent References:
| WO2018225858A1 | 2018-12-13 |
Foreign References:
| JPH06277088A | 1994-10-04 | |||
| JP2004283111A | 2004-10-14 | |||
| JP2010053132A | 2010-03-11 | |||
| JP6940086B1 | 2021-09-22 |
Other References:
SINKUNAS TOMAS ET AL: "Cas3 is a single-stranded DNA nuclease and ATP-dependent helicase in the CRISPR/Cas immune system", EMBO (EUROPEAN MOLECULAR BIOLOGY ORGANIZATION) JOURNAL, vol. 30, no. 7, 1 April 2011 (2011-04-01), pages 1335 - 1342, XP002765626, DOI: 10.1038/emboj.2011.41
KAWAGUCHI YUKO, OKAMOTO, KEN: "3. Gene Delivery and Expression Series. Overexpression of Recombinant Protein in Large-scale Culture Using a Baculovirus-insect Cell Expression System: Based on the Study of Xanthine Oxidoreductase (3)), non-official translation", NIHON IKA DAIGAKU IGAKKAI ZASSHI, vol. 8, no. 1, 1 January 2012 (2012-01-01), pages 26 - 30, XP055965066, ISSN: 1349-8975
HIDEKI YAMAJI: "Production of Useful Substances Using Insect Cells", SEIBUTSU KOGAKU KAISHI = JOURNAL OF THE SOCIETY FOR BIOSCIENCE AND BIOENGINEERING, JAPAN, NIPPON SEIBUTSU KOGAKKAI, SUITA,, JP, vol. 94, no. 2, 1 January 2016 (2016-01-01), JP , pages 76 - 80, XP055965072, ISSN: 0919-3758
JAMIESON A. L. HOWARD, STEPHANE DELMAS, IVANA IVANČIĆ‑BAĆE, EDWARD L. BOLT: "Helicase dissociation and annealing of RNA-DNA hybrids by Escherichia coli Cas3 protein", BIOCHEMICAL JOURNAL, PUBLISHED BY PORTLAND PRESS ON BEHALF OF THE BIOCHEMICAL SOCIETY., vol. 9, no. 1, 1 October 2011 (2011-10-01), pages 4163 - 95, XP055053995, ISSN: 02646021, DOI: 10.1042/BJ20110901
IVANČIĆ-BAĆE IVANA, RADOVČIĆ MARIN, BOČKOR LUKA, HOWARD JAMIESON L., BOLT EDWARD L.: "Cas3 stimulates runaway replication of a ColE1 plasmid in Escherichia coli and antagonises RNaseHI", RNA BIOLOGY, vol. 10, no. 5, 1 May 2013 (2013-05-01), pages 770 - 778, XP055965083, ISSN: 1547-6286, DOI: 10.4161/rna.23876
MAJSEC KRISTINA, BOLT EDWARD L., IVANČIĆ-BAĆE IVANA: "Cas3 is a limiting factor for CRISPR-Cas immunity in Escherichia coli cells lacking H-NS", BMC MICROBIOLOGY, vol. 16, no. 1, 1 December 2016 (2016-12-01), XP055965085, DOI: 10.1186/s12866-016-0643-5
HUO Y. ET AL., NAT. STRUCT. MOL. BIOL., vol. 21, no. 9, September 2014 (2014-09-01), pages 771 - 777
MULEPATI S.BAILEY S., J. BIOL. CHEM., vol. 288, no. 31, 2 August 2013 (2013-08-02), pages 22184 - 22192
ROBERT P. HAYES ET AL., NATURE, vol. 530, 2016, pages 499 - 503
MULEPATI S.BAILEY S., J BIOL CHEM., vol. 288, no. 31, 2 August 2013 (2013-08-02), pages 22184 - 22192
KAWAGUCHI YUKO, OKAMOTO, KEN: "3. Gene Delivery and Expression Series. Overexpression of Recombinant Protein in Large-scale Culture Using a Baculovirus-insect Cell Expression System: Based on the Study of Xanthine Oxidoreductase (3)), non-official translation", NIHON IKA DAIGAKU IGAKKAI ZASSHI, vol. 8, no. 1, 1 January 2012 (2012-01-01), pages 26 - 30, XP055965066, ISSN: 1349-8975
HIDEKI YAMAJI: "Production of Useful Substances Using Insect Cells", SEIBUTSU KOGAKU KAISHI = JOURNAL OF THE SOCIETY FOR BIOSCIENCE AND BIOENGINEERING, JAPAN, NIPPON SEIBUTSU KOGAKKAI, SUITA,, JP, vol. 94, no. 2, 1 January 2016 (2016-01-01), JP , pages 76 - 80, XP055965072, ISSN: 0919-3758
JAMIESON A. L. HOWARD, STEPHANE DELMAS, IVANA IVANČIĆ‑BAĆE, EDWARD L. BOLT: "Helicase dissociation and annealing of RNA-DNA hybrids by Escherichia coli Cas3 protein", BIOCHEMICAL JOURNAL, PUBLISHED BY PORTLAND PRESS ON BEHALF OF THE BIOCHEMICAL SOCIETY., vol. 9, no. 1, 1 October 2011 (2011-10-01), pages 4163 - 95, XP055053995, ISSN: 02646021, DOI: 10.1042/BJ20110901
IVANČIĆ-BAĆE IVANA, RADOVČIĆ MARIN, BOČKOR LUKA, HOWARD JAMIESON L., BOLT EDWARD L.: "Cas3 stimulates runaway replication of a ColE1 plasmid in Escherichia coli and antagonises RNaseHI", RNA BIOLOGY, vol. 10, no. 5, 1 May 2013 (2013-05-01), pages 770 - 778, XP055965083, ISSN: 1547-6286, DOI: 10.4161/rna.23876
MAJSEC KRISTINA, BOLT EDWARD L., IVANČIĆ-BAĆE IVANA: "Cas3 is a limiting factor for CRISPR-Cas immunity in Escherichia coli cells lacking H-NS", BMC MICROBIOLOGY, vol. 16, no. 1, 1 December 2016 (2016-12-01), XP055965085, DOI: 10.1186/s12866-016-0643-5
HUO Y. ET AL., NAT. STRUCT. MOL. BIOL., vol. 21, no. 9, September 2014 (2014-09-01), pages 771 - 777
MULEPATI S.BAILEY S., J. BIOL. CHEM., vol. 288, no. 31, 2 August 2013 (2013-08-02), pages 22184 - 22192
ROBERT P. HAYES ET AL., NATURE, vol. 530, 2016, pages 499 - 503
MULEPATI S.BAILEY S., J BIOL CHEM., vol. 288, no. 31, 2 August 2013 (2013-08-02), pages 22184 - 22192
Attorney, Agent or Firm:
CENTCREST IP ATTORNEYS (JP)
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