First report on CRISPR/Cas9 mediated editing of the eye colour gene, Tryptophan 2, 3-dioxygenase in egg plant shoot and fruit borer Leucinodes orbonalis Guenée (Lepidoptera: Crambidae)

Overuse of synthetic chemicals over a long period of time has not only resulted in control failures but also enormous ecological damage. This necessitates developing eco-friendly, effective alternatives for sustainable pest management. In this regard, potential of CRISPR/Cas9 mediated genome editing to introduce site specific mutations that mostly resulting in loss of function has been successfully demonstrated in wide varieties of organisms including insects. This has opened a new avenue to design and implement futuristic pest management strategies like precision guided sterile insect technique (pgSIT) for an area wide suppression. This requires validation of target genes employing Ribonucleoprotein (RNP) complex containing sgRNA and Cas9 protein before undertaking transgenesis for achieving pgSIT. In the present study, we have for the first time, supporting CRISPR/Cas9 mediated editing of the eye colour gene, Tryptophan 2, 3-dioxygenase of the eggplant shoot and fruit borer, Leucinodes orbonalis using RNP complex. The mutant moths of both sexes exhibited reddish brown eyed phenotype. Having established an RNP mediated editing system in L. orbonalis, the key genes involved in sex determination and spermatogenesis will be validated for developing a pgSIT system for L. orbonalis.