Evaluating Efficiency of Ribo Nucleo Protein (RNP) Delivery Via Microinjection in G0 Embryos of Bactrocera dorsalis (Hendel) for CRISPR/Cas9 Mediated Genome Editing

Bactrocera dorsalis is a serious pest that can cause significant economic losses. CRISPR/Cas9 is a reliable and effective approach for site-directed genome editing that modifies DNA in a stable and heritable manner. This ultimately leads to functional disruption in target gene expression. Despite the multitude of advantages that the system offers, the utility of the method is restricted due to lower mutagenesis efficiency witnessed so far. In the present investigation we optimized the egg laying behavior of tephritids to obtain enough eggs for microinjection at G0 stage. The entire microinjection experiment was planned one hour after egg laying. Consequently, a relatively higher mutagenesis efficiency of 2.72% was witnessed. Therefore, our study was successful in standardising the microinjection process in dipteran insects.