Mapping and CRISPR homology-directed repair of a recessive white eye mutation in Plodia moths

Christa Heryanto; Joseph J Hanly; Anyi Mazo-Vargas; Amruta Tendolkar; Arnaud Martin

doi:10.1016/j.isci.2022.103885

Mapping and CRISPR homology-directed repair of a recessive white eye mutation in Plodia moths

iScience. 2022 Feb 5;25(3):103885. doi: 10.1016/j.isci.2022.103885. eCollection 2022 Mar 18.

Authors

Christa Heryanto¹, Joseph J Hanly¹, Anyi Mazo-Vargas¹, Amruta Tendolkar¹, Arnaud Martin¹

Affiliation

¹ Department of Biological Sciences, The George Washington University, 800 22nd Street NW, Washington, DC 20052, USA.

Abstract

The pantry moth Plodia interpunctella is a worldwide pest of stored food products and a promising laboratory model system for lepidopteran functional genomics. Here we describe efficient methods for precise genome editing in this insect. A spontaneous recessive white-eyed phenotype maps to a frameshift deletion (c.737delC) in the white gene. CRISPR NHEJ mutagenesis of white replicates this phenotype with high rates of somatic biallelic knockout. G₀ individuals with mutant clones on both eyes produced 100% mutant progeny, making white an ideal marker for co-conversion when targeting other genes. CRISPR HDR experiments corrected c.737delC and reverted white eyes to a pigmented state in 37% of G₀ mosaic adults. These repaired alleles showed practical rates of germline transmission in backcrosses, demonstrating the potential of the technique for precise genome editing. Plodia offers a promising avenue for research in this taxon because of its lab-ready features, egg injectability, and editability.

Keywords: Entomology; Genetic engineering; Genomics; Techniques in genetics.