Transcriptomic changes in the pre-blastoderm embryos of the Oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) microinjected with white locus CRISPR/Cas9 RNP complex

CRISPR/Cas9-based genome editing is currently revolutionizing applications in agriculture, human health and other domains. Targeted gene editing employing CRISPR/Cas9 is achieved by delivering the various components viz. guide RNA, traCrRNA and the Cas9 protein in DNA or RNA or RNP format either as a two or three-component system. Among the above, ribonucleoprotein complex (RNP) is more useful in achieving quick, DNA-free editing without the need for nucleic acid integration into the genome of the target species. Employing the RNP complex possesses an advantage over other formats for quick and efficient genome editing. Much research explored the innate immunity against the delivered guide RNA in human cells and plants and the same is lacking in insects. In the present study, the RNP complex (sgRNA+Cas9) was administered via microinjection of embryos (eggs) in the G0 (pre-blastoderm) stage. Transcriptomics was performed using the Illumina Novoseq 6000 platform to understand the prevalence of innate immunity against the delivered guide RNA for the white locus of Bactrocera dorsalis.