Induction of Vermillion in Pyralid moths using CRISPR mutagenesis

Authors: Shirk, Paul; MARTIN, ARNAUD - George Washington University; PATEL, NIPAM - University Of California; SHIRK, BRYCE - Non ARS Employee; Furlong, Richard

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 5/6/2017
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Eye color mutations have been useful markers of genetic activity or alteration in insect genetics. Complementation or disruption of transport or biosynthesis of ommochrome (brown) or pteridine (red) pigments have provided useful targets for transgenic procedures. Tryptophan oxygenase (To) (EC 1.13.1.11) (Drosophila melanogaster vermillion; Ephestia kuehniella a) results in a reddish color eye. The To gene in the Indian Meal moth, Plodia interpunctella, was selected as a convenient gene for visual recognition of mosaic Knock-Outs in the eye color. CRISPR activity was initially confirmed by transfection of Cas9 + To sgRNA into the IAL-PID2 cell line of P. interpunctella. RT-qPCR showed no reduction in To transcript. Microinjection of To directed CRISPR/cas9 into P. interpunctella embryos led to somatic reduction of To transcript and the appearance of the a phenotype in G0 larvae, pupae and adults. The P. interpunctella a phenotype was assessed for germline inheritance. Somatic Knock-Out of the Hox gene Ultrabithorax (Ubx) was also observed as hindwing-to-forewing homeotic transformations in surviving adults. The application of the CRISPR methodology is discussed as a strategic tool for functional genomics in Lepidoptera.