Global gene expression changes induced by knockout of a protease gene cluster in Helicoverpa armigera with CRISPR/Cas9.

Helicoverpa armigera is one of the most serious agricultural insect pests of global importance. It is highly polyphagous and depends on digestive serine proteases to degrade proteins to peptides and to amino acids. H. armigera has evolved adaptive ability to compensate for the inhibition of plant defensive protease inhibitors (PIs) in its diet by overproduction of digestive enzymes. As far as we know, compensation for deletion of serine protease genes has not yet been studied in any herbivorous insect. In this study, we used CRISPR/Cas9 to knock out a cluster of 18 trypsin-like genes in H. armigera. Compared with the wild type SCD strain, activities of the total proteases, trypsins and chymotrypsins were not significantly changed in the gene cluster knockout strain (Tryp-KO). RNA-seq data showed 1492 upregulated and 461 downregulated DEGs in Try-KO. GO function classification and KEGG pathway analyses revealed these differentially expressed genes were enriched for terms related to binding, catalytic activity, metabolic process and signal transduction. In regard to serine protease genes, 35 were upregulated and 12 downregulated in Tryp-KO strain. Our study indicated that H. armigera can compensate for the deleted protease genes by overexpression of other trypsin and chymotrypsin genes in order to maintain its genetic and metabolic robustness. It also suggests that genetic perturbations created by genome editing tools can induce global gene expression changes.