APPLICATION OF CRISPR/CAS12A GENE EDITING SYSTEM IN GENE EDITING OF PHYSCOMITRELLA PATENS

Some embodiments of the disclosure provide an application method of a CRISPR/Cas12a gene editing system in the gene editing of Physcomitrella patens (P. patens). According to an embodiment， the application method includes the following steps: 1) constructing a Cas12a protease expression vector by ligating a Cas12a-protease-encoding nucleotide sequence with nuclear localization signals at both ends to a plasmid pAct-Cas9 and initiating expression by a pActin promoter; 2) constructing a gRNA expression vector by ligating a gRNA to a plasmid pU6-sgRNA， and initiating expression by a PpU6 promoter; and 3) transforming P. patens by using the Cas12a protease expression vector， the gRNA expression vector， and the plasmid for resistance expression obtain a mutant plant through screening for resistance.