The invention discloses construction of an efficient
gene editing
system based on
CRISPR-Cpf1, and belongs to the technical field of
gene engineering. The method comprises the following steps: firstly, representing a Cas
protein Cpf1 from F.novicida, and integrating FnCpf1 to a lacA site of a
bacillus subtilis genome; the crRNA targeting different genes is placed at the downstream of a strong constitutive
promoter Pveg from
bacillus subtilis and is used for high-intensity expression of the corresponding crRNA. By selecting crRNA of different target genes, the
genome editing method is verified. A
bacterial colony PCR (
Polymerase Chain Reaction) result shows that the sacA
gene, the ligDV gene and the pps
gene cluster can be efficiently knocked out. Meanwhile, in order to verify the knock-in efficiency of the
system, the exogenous gene sfGFP is efficiently knocked out to the sacA site. In order to further verify the
polygene knockout efficiency of the
system, sacA and aprE are selected as targets, and a
bacterial colony PCR result shows high knockout efficiency.