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Research Project: IPM Methods for Insect Pests of Orchard Crops

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Title: APC-assisted-CRISPR-Cas9 delivery into nymphs and adults for heritable gene editing (Hemiptera)

Author
item Hunter, Wayne
item Gonzalez, Maria
item TOMICH, JOHN - Kansas State University

Submitted to: American Society for Biochemistry and Molecular Biology Annual Meeting
Publication Type: Proceedings
Publication Acceptance Date: 12/7/2019
Publication Date: 3/6/2019
Citation: Hunter, W.B., Gonzalez, M.T., Tomich, J. 2019. BAPC-assisted-CRISPR-Cas9 delivery into nymphs and adults for heritable gene editing (Hemiptera). In: Proceedings of American Society for Biochemistry and Molecular Biology Annual Meeting. 626.2.

Interpretive Summary: A new method enable the disabling of specific insect genes in Diaphorina citri, Asian citrus psyllid which will be passed to subsequent offspring. Reduced fitness can also reduce psyllid populations. The method could also be used to turn the psyllid vector, into a non-vector by blocking gene critical for pathogen transmission. These efforts may reduce citrus greening disease, huanglongbing, and are key development to have a sustainable citrus industry.

Technical Abstract: A new method for heritable gene editing in insects, specifically Hemiptera, used Branched Amphiphilic Peptide Capsules, BAPC, to delivery guide Ribonucleic acids and other components into insect ovarian cells resulting in heritable gene editing. The method permits injection of the nymph and adult stages of insects, bypassing the need for embryonic injections. First heritable gene knockouts in Asian citrus psyllid- Diaphorina citri. Second generation psyllids had slower development, shorter adult lifespan, and reduced fecundity. The method permits altering psyllid vectors into non-vectors, stopping Liberibacter transmission. Slower development time provides parasitoids and predators more time, 2-3 weeks longer, to attack nymphal stages reducing populations further. The BAPC-assisted delivery system advances gene-editing efforts across all hemipterans and other insects by permitting the use of nymphs and adults. [citrusgreening.org]. USDA-NIFA 2014-70016-23028. Developing an infrastructure and product test pipeline to deliver novel therapies for citrus greening disease.