CRISPR/Cas9 mediated knockout of the abdominal-A homeotic gene in fall armyworm moth (Spodoptera frugiperda)

The fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), is an important pest of maize in the Americas and has recently been introduced into Africa. Fall armyworm populations have developed resistance to control strategies that depend on insecticides and transgenic plants expressing Bacillus thu...

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Veröffentlicht in:PloS one 2018-12, Vol.13 (12), p.e0208647-e0208647
Hauptverfasser: Wu, Ke, Shirk, Paul D, Taylor, Caitlin E, Furlong, Richard B, Shirk, Bryce D, Pinheiro, Daniele H, Siegfried, Blair D
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container_issue 12
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container_title PloS one
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creator Wu, Ke
Shirk, Paul D
Taylor, Caitlin E
Furlong, Richard B
Shirk, Bryce D
Pinheiro, Daniele H
Siegfried, Blair D
description The fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), is an important pest of maize in the Americas and has recently been introduced into Africa. Fall armyworm populations have developed resistance to control strategies that depend on insecticides and transgenic plants expressing Bacillus thuringiensis toxins. The study of various resistance mechanisms at the molecular level and the development novel control strategies have been hampered by a lack of functional genomic tools such as gene editing in this pest. In the current study, we explored the possibility of using the CRISPR/Cas9 system to modify the genome of FAW. We first identified and characterized the abdominal-A (Sfabd-A) gene of FAW. Sfabd-A single guide RNA (sgRNA) and Cas9 protein were then injected into 244 embryos of FAW. Sixty-two embryos injected with Sfabd-A sgRNA hatched. Of these hatched embryos, twelve developed into larvae that displayed typical aba-A mutant phenotypes such as fused segments. Of the twelve mutant larvae, three and five eventually developed into female and male moths, respectively. Most mutant moths were sterile, and one female produced a few unviable eggs when it was outcrossed to a wild-type male. Genotyping of 20 unhatched Sfabd-A sgRNA-injected embryos and 42 moths that developed from Sfabd-A sgRNA-injected embryos showed that 100% of the unhatched embryos and 50% of the moths contained indel mutations at the Sfabd-A genomic locus near the guide RNA target site. These results suggest that the CRISPR/Cas9 system is highly efficient in editing FAW genome. Importantly, this gene editing technology can be used to validate gene function to facilitate an understanding of the resistance mechanism and lead to the development of novel pest management approaches.
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Smith), is an important pest of maize in the Americas and has recently been introduced into Africa. Fall armyworm populations have developed resistance to control strategies that depend on insecticides and transgenic plants expressing Bacillus thuringiensis toxins. The study of various resistance mechanisms at the molecular level and the development novel control strategies have been hampered by a lack of functional genomic tools such as gene editing in this pest. In the current study, we explored the possibility of using the CRISPR/Cas9 system to modify the genome of FAW. We first identified and characterized the abdominal-A (Sfabd-A) gene of FAW. Sfabd-A single guide RNA (sgRNA) and Cas9 protein were then injected into 244 embryos of FAW. Sixty-two embryos injected with Sfabd-A sgRNA hatched. Of these hatched embryos, twelve developed into larvae that displayed typical aba-A mutant phenotypes such as fused segments. 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subjects Abdomen
Analysis
Animals
Base Sequence
Biology and Life Sciences
Butterflies & moths
Corn
CRISPR
CRISPR-Cas Systems
Deoxyribonucleic acid
DNA
Eggs
Embryos
Engineering and Technology
Female
Gene Knockdown Techniques
Genes
Genetic engineering
Genetic modification
Genetically modified plants
Genome editing
Genomes
Genomics
Genotyping
Insect Proteins - genetics
Insect Proteins - metabolism
Insecticide resistance
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Insects
Larvae
Male
Mutagenesis, Site-Directed
Mutation
Novels
Pest control
Pest Control, Biological
Pests
Phenotype
Phenotypes
Phylogeny
Proteins
Research and Analysis Methods
Ribonucleic acid
RNA
Spodoptera - anatomy & histology
Spodoptera - genetics
Spodoptera - metabolism
Spodoptera frugiperda
Technology
Toxins
Transgenic plants
title CRISPR/Cas9 mediated knockout of the abdominal-A homeotic gene in fall armyworm moth (Spodoptera frugiperda)
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