The invention relates to the technical field of
gene engineering, in particular to a method for rapidly and economically carrying out
gene function analysis on lepidoptera insects and application. The method comprises the following steps: designing four different sgRNAs by aiming at an
exon region of a
target gene, and constructing a sgRNAs synthetic primer mixing
pool; 4 different sgRNA-
DNA templates are synthesized at the same time, 4 different sgRNAs are further synthesized at the same time, and a sgRNAs mixed
pool is formed; the sgRNAs-Cas9
protein and Cas9
protein are prepared into an sgRNAs-Cas9 compound, and the sgRNAs-Cas9 compound is microinjected into newly laid eggs; after the injected eggs develop into larvae,
genome DNA is extracted, sgRNAs targeted areas are amplified, the
mutation rate of target genes is determined through T7E1
enzyme digestion, and
phenotype analysis is conducted on the larvae. According to the method, the
mutation rate of the
target gene is increased through redundant targeting of the four sgRNAs, so that
functional analysis of the
target gene is quickly realized in the G0 generation, and the research period of the
gene function of the lepidoptera insects is greatly shortened. Besides, four different sgRNAs are synthesized at the same time by constructing a primer mixing
pool of a plurality of sgRNAs, so that the economic cost for synthesizing the sgRNAs is greatly reduced.