Title:
NUCLEIC ACID CONSTRUCT BASED ON CRE-LOXP AND CRISPR AND USE THEREOF
Document Type and Number:
WIPO Patent Application WO/2023/226856
Kind Code:
A1
Abstract:
Provided are a nucleic acid construct based on a Cre-LoxP recombination system and a CRISPR gene editing system and use thereof. The Cre-LoxP recombination system comprises a Cre enzyme and a LoxP nucleic acid combination. The LoxP nucleic acid combination comprises TATA-Lox71 and TATA-LoxTC9 sequences, which can only be recombined once under the catalysis of the Cre enzyme. The nucleic acid construct carries an inert "filling sequence" with a certain length. The nucleic acid construct can express a plurality of sgRNAs in a low bias manner in vivo, but a same cell can only express one sgRNA, thereby efficiently generating a genetic chimera, and use of the latter comprises accurate and sensitive in-situ CRISPR gene screening and rapid and low-cost preparation of a single-gene knockout strain.
Inventors:
CHI TIAN (CN)
LIU BO (CN)
JING ZHENYU (CN)
ZHANG XIAOMING (CN)
MAO SHAOSHUAI (CN)
CHEN YUXIN (CN)
LIU BO (CN)
JING ZHENYU (CN)
ZHANG XIAOMING (CN)
MAO SHAOSHUAI (CN)
CHEN YUXIN (CN)
Application Number:
PCT/CN2023/094885
Publication Date:
November 30, 2023
Filing Date:
May 17, 2023
Export Citation:
Assignee:
UNIV SHANGHAI TECH (CN)
International Classes:
C12N15/85; A01K67/027; C12N5/10
Domestic Patent References:
| WO2017048995A1 | 2017-03-23 |
Foreign References:
| CN1283233A | 2001-02-07 | |||
| US20190085325A1 | 2019-03-21 | |||
| CN106637421A | 2017-05-10 | |||
| CN108103586A | 2018-06-01 |
Other References:
LIU BO, JING ZHENGYU, ZHANG XIAOMING, CHEN YUXIN, MAO SHAOSHUAI, KAUNDAL RAVINDER, ZOU YAN, WEI GE, ZANG YING, WANG XINXIN, LIN WE: "Large-scale multiplexed mosaic CRISPR perturbation in the whole organism", CELL, ELSEVIER, AMSTERDAM NL, vol. 185, no. 16, 1 August 2022 (2022-08-01), Amsterdam NL , pages 3008 - 3024.e16, XP093111735, ISSN: 0092-8674, DOI: 10.1016/j.cell.2022.06.039
CHEN YUXIN, MAO SHAOSHUAI, LIU BO, JING ZHENGYU, ZANG YING, XIA JING, SUN JIANLONG, CHI TIAN: "Novel mosaic mice with diverse applications", BIORXIV, 24 March 2020 (2020-03-24), XP093111737, Retrieved from the Internet [retrieved on 20231213], DOI: 10.1101/2020.03.21.001388
PONTES-QUERO SAMUEL; HEREDIA LUIS; CASQUERO-GARCíA VERóNICA; FERNáNDEZ-CHACóN MACARENA; LUO WEN; HERMOSO ANA; : "Dual ifgMosaic: A Versatile Method for Multispectral and Combinatorial Mosaic Gene-Function Analysis", CELL, ELSEVIER, AMSTERDAM NL, vol. 170, no. 4, 10 August 2017 (2017-08-10), Amsterdam NL , pages 800, XP085153698, ISSN: 0092-8674, DOI: 10.1016/j.cell.2017.07.031
TAKURO HORII, SUMIYO MORITA, MIKA KIMURA, NAOMI TERAWAKI, MIHIRO SHIBUTANI, IZUHO HATADA: "Efficient generation of conditional knockout mice via sequential introduction of lox sites", SCIENTIFIC REPORTS, vol. 7, no. 1, 1 December 2017 (2017-12-01), XP055516908, DOI: 10.1038/s41598-017-08496-8
WANG CHAOXUAN, SUN HANG: "Progress in Gene Knockout Mice", CHINESE JOURNAL OF BIOTECHNOLOGY, ZHONGGUO KEXUEYUAN WEISHENGWU YANJIUSUO, CHINESE ACADEMY OF SCIENCES, INSTITUTE OF MICROBIOLOGY, CN, vol. 35, no. 5, 13 March 2019 (2019-03-13), CN , pages 784 - 794, XP093111789, ISSN: 1000-3061, DOI: 10.13345/j.cjb.180417
CHEN YUXIN, MAO SHAOSHUAI, LIU BO, JING ZHENGYU, ZANG YING, XIA JING, SUN JIANLONG, CHI TIAN: "Novel mosaic mice with diverse applications", BIORXIV, 24 March 2020 (2020-03-24), XP093111737, Retrieved from the Internet
PONTES-QUERO SAMUEL; HEREDIA LUIS; CASQUERO-GARCíA VERóNICA; FERNáNDEZ-CHACóN MACARENA; LUO WEN; HERMOSO ANA; : "Dual ifgMosaic: A Versatile Method for Multispectral and Combinatorial Mosaic Gene-Function Analysis", CELL, ELSEVIER, AMSTERDAM NL, vol. 170, no. 4, 10 August 2017 (2017-08-10), Amsterdam NL , pages 800, XP085153698, ISSN: 0092-8674, DOI: 10.1016/j.cell.2017.07.031
TAKURO HORII, SUMIYO MORITA, MIKA KIMURA, NAOMI TERAWAKI, MIHIRO SHIBUTANI, IZUHO HATADA: "Efficient generation of conditional knockout mice via sequential introduction of lox sites", SCIENTIFIC REPORTS, vol. 7, no. 1, 1 December 2017 (2017-12-01), XP055516908, DOI: 10.1038/s41598-017-08496-8
WANG CHAOXUAN, SUN HANG: "Progress in Gene Knockout Mice", CHINESE JOURNAL OF BIOTECHNOLOGY, ZHONGGUO KEXUEYUAN WEISHENGWU YANJIUSUO, CHINESE ACADEMY OF SCIENCES, INSTITUTE OF MICROBIOLOGY, CN, vol. 35, no. 5, 13 March 2019 (2019-03-13), CN , pages 784 - 794, XP093111789, ISSN: 1000-3061, DOI: 10.13345/j.cjb.180417
Attorney, Agent or Firm:
SHANGHAI BESHINING LAW OFFICE (CN)
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