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Inducible inheritance recombinase system CrexER

A technology of recombinase and recombination site, applied in the direction of recombinant DNA technology, other methods of inserting foreign genetic materials, isomerase, etc., can solve the problems of Cre expression specificity or precision not high enough, experimental result interpretation inaccurate, etc.

Active Publication Date: 2018-05-25
CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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AI Technical Summary

Problems solved by technology

Although the traditional genetic manipulation technology based on Cre-loxP homologous recombination has very powerful functions, when the expression specificity or precision of Cre is not high enough, it will cause inaccurate interpretation of many experimental results and even cause great controversy

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  • Inducible inheritance recombinase system CrexER
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  • Inducible inheritance recombinase system CrexER

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Embodiment Construction

[0031] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described in the following (such as embodiments) can be combined with each other to form a preferred technical solution.

[0032] This article provides an inducible homologous recombination enzyme CrexER, which uses the prior occurrence of Dre-rox homologous recombination to induce the subsequent occurrence of Cre-loxP homologous recombination, and realizes the intersection of A-Dre and B-Cre labeled cells (A + B + ) for targeted manipulation.

[0033] Specifically, the homologous recombinase CrexER herein contains Cre recombinase, ER and rox sequences.

[0034]The Cre recombinase suitable for this article may be the Cre recombinase well known in the art. The full-length sequence of its gene coding region is 1029 bp (EMBL database accession number X03453), encoding a 38 kDa monomeric protein cons...

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Abstract

The invention relates to an inducible inheritance recombinase system CrexER. The CrexER is a fusion protein; the fusion protein is a protein formed by fusing recombinase Cre, estrogen receptors ER andrecombination sites rox specifically recognized by recombinase Dre. The invention also provides a coding sequence of the fusion protein, a nucleic acid construct containing the coding sequence, a host cell and the like. The ahead occurrence of Dre-rox homologous recombination is used for inducing the subsequent occurrence of Cre-loxP homologous recombination; the targeted operation of the A-Dre and B-Cre mark cell intersection (A+B+) is realized. The application range of the inheritance targeted operation is greatly expanded; a precious strategy is provided for the precise inheritance targeted operation and biomedical research.

Description

technical field [0001] The present invention relates to an inducible genetic recombinase system CrexER. Background technique [0002] In the past 10 years, genetic manipulation techniques used in mouse models have greatly advanced the understanding of many human diseases, and the most widely used genetic manipulation tools are mainly based on the Cre-loxP homologous recombination system. So far, thousands of genetic tool mice containing Cre or loxP sites have been used to address various questions about cell lineage, organ development, tissue regeneration, and disease. The accuracy of this gene targeting system largely depends on the specificity of Cre expression, usually Cre is expressed under the drive of a gene promoter. Although the traditional genetic manipulation technology based on Cre-loxP homologous recombination has very powerful functions, when the expression specificity or precision of Cre is not high enough, it will cause inaccurate interpretation of many exper...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/90
CPCC07K14/721C07K2319/00C12N9/90C12N15/902
Inventor 周斌蒲文娟何玲娟
Owner CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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