Poster Display
Physiology, Biochemistry, and Toxicology
Jennifer Lynn Stout
USDA-ARS
Wapato, Washington
William Walker
Research Geneticist
USDA-ARS
Wapato, Washington
The codling moth, Cydia pomonella (L.), (Lepidoptera:Tortricidae), is a global pest of apple, pear and walnut. CRISPR/Cas9 gene editing serves as an excellent way to study the biological effects of individual genes, in vivo, of non-model organisms, and has previously been demonstrated to function efficiently in codling moth. However, challenges remain in tracking the CRISPR-edited genotypes without utilization of good phenotypic markers. Producing a phenotypic white eye moth by knocking out the white transporter gene, would provide a convenient visual marker to facilitate targeting other genes for CRISPR editing. When a homozygous white eye strain of moths is created, CRISPR could be used to knock in a wild-type copy of the white gene into the exon of any target gene of interest, thereby rescuing the white eye phenotype. This would effectively result in a normal wild type looking moth that also has a knocked-out target gene of interest. For this study, we have targeted the white gene in codling moth for CRISPR/CAS9 editing. By first targeting the white gene, (KNOCK OUT) we have generated visual marker strains that can be used later to ensure our gene of interest has been edited (KNOCK IN/KNOCK OUT). We report on the phenotypic effects of the white mutant strains as well as the genomic dynamics of CRISPR editing at this gene locus, with respect to genomic DNA and mRNA transcripts, and discuss downstream implications in genome-edited codling moth.