CRISPR/Cpf1 gene editing system, as well as construction method of CRISPR/Cpf1 gene editing system and application of CRISPR/Cpf1 gene editing system in gibberella

A gene editing and construction method technology, applied in the field of genetic engineering, can solve the problems of low efficiency of multi-gene editing, speed up the production of bacterial strains, and low efficiency of integration technology, and achieve the effect of simple system, easy repair, and favorable plasmid transformation
CN113584033AActive Publication Date: 2021-11-02NANJING NORMAL UNIVERSITY
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
NANJING NORMAL UNIVERSITY
Publication Date
2021-11-02

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Abstract

The invention discloses a CRISPR (clustered regularly interspaced short palindromic repeats) / Cpf1 gene editing system as well as a construction method and the application thereof in gibberella. The system is a skeleton plasmid pCpf1Ff-DR, and comprises an Fncpf1 gene, a functional crRNA gene, a promoter pGPD, a promoter pTRPC, a strong RNA polymerase III type promoter 5srRNA and a hygromycin selection marker hph. The system is high in efficiency, strong in universality, easy to operate and capable of being rapidly applied to filamentous fungus gibberella, and the gene editing system can be used for achieving knockout of the bicarpin gene cluster as long as 17kb in the gibberella. According to the method, large-fragment gene knockout is achieved in gibberellin for the first time, it is found for the first time that knockout of the bicarpin gene cluster is beneficial to increase of the yield of gibberellin, and the method is expected to be further used for transforming gibberellin industrial bacteria, shortening the time of bacterial strain upgrading and providing powerful guarantee for increasing the yield of gibberellin and increasing economic benefits.
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Description

technical field

[0001] The invention belongs to the field of genetic engineering, and in particular relates to a CRISPR / Cpf1 gene editing system and its construction method and application in Gibberella. Background technique

[0002] It is of great significance to perform gene editing in filamentous fungi such as gibberellins. Filamentous fungi are a group of microorganisms that play an important role in protein secretion, production of active secondary metabolites, and environmental pollution control. Both basic and applied research are highly dependent on gene editing platforms. Filamentous fungi are of great value in the commercial production of enzymes, heterologous proteins, a wide range of primary and secondary metabolites, and biofuels. One of the ways to improve the commercial potential of fungi is to improve strains through genetic engineering. However, the physiological characteristics of filamentous fungi, such as apical growth, heterokaryosis, low homologous rec...

Claims

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