Toward a CRISPR-Cas9-based gene drive in the diamondback moth Plutella xylostella

Xu, Xuejiao, Harvey-Samuel, Tim, Siddiqui, Hamid Anees, Ang, Joshua Xin De, Anderson, Michelle Ellis, Reitmayer, Christine M., Lovett, Erica, Leftwich, Philip T. ORCID:, You, Minsheng and Alphey, Luke (2022) Toward a CRISPR-Cas9-based gene drive in the diamondback moth Plutella xylostella. The CRISPR Journal, 5 (2). pp. 224-236. ISSN 2573-1599

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Promising to provide powerful genetic control tools, gene drives have been constructed in multiple dipteran insects, yeast, and mice for the purposes of population elimination or modification. However, it remains unclear whether these techniques can be applied to lepidopterans. Here, we used endogenous regulatory elements to drive Cas9 and single guide RNA (sgRNA) expression in the diamondback moth (DBM), Plutella xylostella, and test the first split gene drive system in a lepidopteran. The DBM is an economically important global agriculture pest of cruciferous crops and has developed severe resistance to various insecticides, making it a prime candidate for such novel control strategy development. A very high level of somatic editing was observed in Cas9/sgRNA transheterozygotes, although no significant homing was revealed in the subsequent generation. Although heritable Cas9-medated germline cleavage as well as maternal and paternal Cas9 deposition were observed, rates were far lower than for somatic cleavage events, indicating robust somatic but limited germline activity of Cas9/sgRNA under the control of selected regulatory elements. Our results provide valuable experience, paving the way for future construction of gene drives or other Cas9-based genetic control strategies in DBM and other lepidopterans.

Item Type: Article
Additional Information: Funding Information: This work was supported by European Union H2020 Grant nEUROSTRESSPEP (634361). In addition, THS and LA were respectively supported by a Impact Acceleration Account grant (BB/S506680/1), and three core fundings [BBS/E/I/00007033, BBS/E/I/00007038 and BBS/E/I/00007039] from the UK Biotechnology and Biological Sciences Research Council. XX was supported by a CSC Scholarship from the Chinese Government.
Faculty \ School: Faculty of Science > School of Biological Sciences
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Depositing User: LivePure Connector
Date Deposited: 23 Mar 2022 14:30
Last Modified: 26 Oct 2022 00:03
DOI: 10.1089/crispr.2021.0129


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