Whole-genome random mutation method based on CRISPR-Cas system and application of whole-genome random mutation method

A whole-genome, random mutation technology, applied in the field of bioengineering, can solve the problems of destroying the fidelity of genome replication, poor stability of mutants, single mutation type, etc., reaching single-cell mutation sites with diverse mutation types and high mutation rate , the effect of simple operation
CN112680450APending Publication Date: 2021-04-20EAST CHINA UNIV OF SCI & TECH
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
EAST CHINA UNIV OF SCI & TECH
Publication Date
2021-04-20

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Abstract

The invention discloses a whole-genome random mutation method based on a CRISPR-Cas system and application of the whole-genome random mutation method. The whole-genome random mutation method is based on the CRISPR-Cas system, through the guidance of a random gRNA library, the whole genome of a living body is subjected to scattered bullet type random cutting, and random mutation on the whole genome scale is triggered under the non-fidelity type DNA repair effect. The whole-genome random mutation method can be used for directed evolution and iterative evolution, saccharomyces cerevisiae is used for producing beta-carotene to serve as a verification model, seven rounds of iterative mutation screening are completed within two months, and an evolved strain with the yield increased to 10.5 times is obtained. The omics analysis shows that about 122 mutations can be introduced into each operation on average, the transcription level of about 50% of genes is remarkably changed, it shows that deep remodeling occurs in saccharomyces metabolism through mutation evolution, and experiments prove that the method is a simple and controllable new genome variation technology.
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Description

technical field

[0001] The invention belongs to the technical field of bioengineering, and more specifically relates to a random mutation method of the whole genome based on a CRISPR-Cas system and its application. Background technique

[0002] Simulating natural evolution and rapid evolution of living organisms in the laboratory is an important means to study complex metabolic traits of cells, develop new functions of cells and realize biological breeding. The idea of ​​laboratory evolution has a long history. Physical or chemical "mutation" as a means of laboratory evolution has appeared as early as the 1920s. After nearly a hundred years of development, a complete technical and theoretical system has been formed, which actively promotes The rapid development of biological industries such as antibiotics, organic acids, vitamins, steroid pharmaceuticals, modern brewing, enzyme preparations, and crop breeding. Today, mutagenesis is still an important means of breeding in ma...

Claims

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