文章基本信息

标题：Mapping and CRISPR homology-directed repair of a recessive white eye mutation in Plodia moths
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作者：Christa Heryanto ; Joseph J. Hanly ; Anyi Mazo-Vargas 等
期刊名称：iScience
印刷版ISSN：2589-0042
出版年度：2022
卷号：25
期号：3
页码：1-20
DOI：10.1016/j.isci.2022.103885
语种：English
出版社：Elsevier
摘要：SummaryThe pantry mothPlodia interpunctellais 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 thewhitegene. CRISPR NHEJ mutagenesis ofwhitereplicates this phenotype with high rates of somatic biallelic knockout. G0individuals with mutant clones on both eyes produced 100% mutant progeny, makingwhitean ideal marker for co-conversion when targeting other genes. CRISPR HDR experiments correctedc.737delCand reverted white eyes to a pigmented state in 37% of G0mosaic adults. These repaired alleles showed practical rates of germline transmission in backcrosses, demonstrating the potential of the technique for precise genome editing.Plodiaoffers a promising avenue for research in this taxon because of its lab-ready features, egg injectability, and editability.Graphical abstractDisplay OmittedHighlights•Plodiapantry moths are an emerging model organism for functional genomics in Lepidoptera•Spontaneous and CRISPR-inducedwhitemutations yield recessive-white eye phenotypes•CRISPR HDR repair with ssODN donor result in practical rates of base editing•We provide optimized protocols forPlodiahandling and genome editingEntomology; Genetic engineering; Genomics; Techniques in genetics