Application of CRISPR/Cas9 Genome Editing Technology in Filamentous Fungi
There are many kinds of secondary metabolites of filamentous fungi,many of which have important bioactive substances and medicinal components,as well as some toxins,so it is very necessary to carry out the study of gene editing.The system can be used to carry out point mutation,insertion or deletion of its genome to a-chieve gene knockout,but the cell wall of filamentous fungi is thick and the composition is complex,so it is diffi-cult to introduce foreign genes.It is very important to develop an efficient CRISPR/Cas9 genome editing technol-ogy.By using the method of literature research,this paper introduces the principle and method of CRISPR/Cas9 genome editing technology and its research and application in filamentous fungi,and analyzes the effects of the optimization of components and presentation system of CRISPR/Cas9 system on gene editing efficiency in fila-mentous fungi.Among them,the optimization of Cas9 nuclease and sgRNA expression,the improvement of pro-moter,PAM sequence and genome editing system have effectively improved the editing efficiency of CRISPR/Cas9 system.In the future,a series of general fungal gene editing systems can be developed,such as no PAM se-quence requirements,gene transcriptional regulation,directional insertion and knockout of ultra-long target frag-ments,so as to realize the full application of fungal resources in the fields of industry,agriculture and medicine.
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