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Title: Efficient CRISPR / Cas9 ‐mediated genome editing in the European corn borer, Ostrinia nubilalis
Abstract The European corn borer (Ostrinia nubilalis) is an agricultural pest and burgeoning model for research on speciation, seasonal adaptation and insect resistance management. Although previous work inO. nubilalishas identified genes associated with differences in life cycle, reproduction, and resistance toBttoxins, the general lack of a robust gene‐editing protocol forO. nubilalishas been a barrier to functional validation of candidate genes. Here, we demonstrate an efficient and practical methodology for heritable gene mutagenesis inO. nubilalisusing the CRISPR/Cas9 genome editing system. Precise loss‐of‐function (LOF) mutations were generated at two circadian clock genes,period(per) andpigment‐dispersing factor receptor(pdfr), and a developmental gene,prothoracicotropic hormone(ptth). Precluding the need for a visible genetic marker, gene‐editing efficiency remained high across different single guide RNAs (sgRNA) and germline transmission of mutations to F1offspring approached 100%. When single or dual sgRNAs were injected at a high concentration, gene‐specific phenotypic differences in behaviour and development were identified in F0mutants. Specifically, F0gene mutants demonstrated that PER, but not PDFR, is essential for normal timing of eclosion. PTTH F0mutants were significantly heavier and exhibited a higher incidence of diapause. This work will accelerate future studies of gene function inO. nubilalisand facilitate the development of similar screens in other Lepidopteran and non‐model insects.  more » « less
Award ID(s):
2416175
PAR ID:
10583539
Author(s) / Creator(s):
; ; ;
Publisher / Repository:
John Wiley & Sons
Date Published:
Journal Name:
Insect Molecular Biology
Volume:
34
Issue:
1
ISSN:
0962-1075
Page Range / eLocation ID:
174 to 184
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
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