Title:
CHLOROPLAST GENOME EDITING METHOD
Document Type and Number:
WIPO Patent Application WO/2018/103686
Kind Code:
A1
Abstract:
Provided is a chloroplast genome editing method. In particular, provided are a nucleic acid construct, a vector, or a vector combination for plant genome site-specific editing based on a CRISPR technology, and a plant genome site-specific editing method. The nucleic acid construct comprises a nucleic acid construct of formula I and/or a nucleic acid construct of formula II, the nucleic acid construct of formula I comprises a chloroplast targeting signal peptide-nuclease expression cassette, and the nucleic acid construct of formula II comprises an ncRNA-sgRNA expression cassette. With this method, a nuclease and a corresponding sgRNA can be introduced into a chloroplast, so that gene knockout or homologous recombination and targeted insertion of a foreign segment can be carried out simply and efficiently at a predetermined chloroplast genomic site. This method can be used to improve a trait of a crop from the chloroplast genome level.
Inventors:
ZHU JIANKANG (CN)
WANG DONG (CN)
HUA KAI (CN)
LIU ZHIHONG (CN)
WANG DONG (CN)
HUA KAI (CN)
LIU ZHIHONG (CN)
Application Number:
PCT/CN2017/114962
Publication Date:
June 14, 2018
Filing Date:
December 07, 2017
Export Citation:
Assignee:
SHANGHAI INST BIOLOGICAL SCIENCES CAS (CN)
International Classes:
C12N15/11; C12N15/09; C12N15/63; C12N15/66
Foreign References:
| CN105602935A | 2016-05-25 | |||
| CN103981215A | 2014-08-13 | |||
| CN103981216A | 2014-08-13 |
Other References:
FRANCIS QUÉTIER: "The CRISPR-Cas9 technology: Closer to the ultimate toolkit for targeted genome editing", PLANT SCIENCE, vol. 242, 8 September 2015 (2015-09-08), pages 65 - 76, XP055613051
VEJNAR, C.E.: "Optimization Strategies for the CRISPR -Cas9 Genome-Edi- ting System", COLD SPRING HARBOR PROTOCOLS, 3 October 2016 (2016-10-03)
CHO, S.W.: "Targeted genome engineering in human cells with the Cas9 RNA- guided endonuclease", NATURE BIOTECHNOLOGY, vol. 31, no. 3, 29 January 2013 (2013-01-29), pages 230 - 232, XP002699850
JINEK, M.: "RNA-programmed genome editing in human cells", ELIFE, vol. 2, 29 January 2013 (2013-01-29), pages e00471, XP002699851
MALI, P.: "RNA-Guided Human Genome Engineering via Cas9", SCIENCE, vol. 339, 15 February 2013 (2013-02-15), pages 823 - 826, XP055469277
CONG, LE: "Multiplex Genome Engineering Using CRISPR/Cas Systems", SCIENCE, vol. 339, 15 February 2013 (2013-02-15), pages 819 - 823, XP055400719
VEJNAR, C.E.: "Optimization Strategies for the CRISPR -Cas9 Genome-Edi- ting System", COLD SPRING HARBOR PROTOCOLS, 3 October 2016 (2016-10-03)
CHO, S.W.: "Targeted genome engineering in human cells with the Cas9 RNA- guided endonuclease", NATURE BIOTECHNOLOGY, vol. 31, no. 3, 29 January 2013 (2013-01-29), pages 230 - 232, XP002699850
JINEK, M.: "RNA-programmed genome editing in human cells", ELIFE, vol. 2, 29 January 2013 (2013-01-29), pages e00471, XP002699851
MALI, P.: "RNA-Guided Human Genome Engineering via Cas9", SCIENCE, vol. 339, 15 February 2013 (2013-02-15), pages 823 - 826, XP055469277
CONG, LE: "Multiplex Genome Engineering Using CRISPR/Cas Systems", SCIENCE, vol. 339, 15 February 2013 (2013-02-15), pages 819 - 823, XP055400719
Attorney, Agent or Firm:
XU & PARTNERS,LLC. (CN)
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