CRISPR/Cas9-mediated knockout of two eye pigmentation genes in the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae)
Autor: | Hao-Hao Chen, Jin-Li Zhang, Sheng-Jie Fu, Xiao-Bo Yuan, Wen-Hua Xue, Nan Xu, Hai-Jun Xu, Chuan-Xi Zhang |
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Rok vydání: | 2018 |
Předmět: |
Male
0301 basic medicine Biochemistry Hemiptera Gene Knockout Techniques 03 medical and health sciences Animals CRISPR Compound Eye Arthropod Molecular Biology Gene Genetics Base Sequence Eye Color biology Pigmentation Cas9 biology.organism_classification Eye pigmentation White (mutation) 030104 developmental biology Insect Science Insect Proteins Female Brown planthopper CRISPR-Cas Systems Delphacidae Functional genomics |
Zdroj: | Insect Biochemistry and Molecular Biology. 93:19-26 |
ISSN: | 0965-1748 |
Popis: | The brown planthopper Nilaparvata lugens is one of the most destructive insect pests in Asia, demonstrating high fertility and causing huge crop losses by sucking sap of rice as well as transmitting viruses. However, functional genomic studies on N. lugens are seriously constrained by lack of genetic tools. Here, we employed two eye pigmentation genes to generate germ-line mutations in N. lugens using the CRISPR/Cas9 (clustered regularly interspaced palindromic repeats/CRISPR-associated) system. We showed that injection of single guide RNA of the cinnabar gene of N. lugens (Nl-cn ) into pre-blastoderm eggs induced insertion and deletion (indels) in the founder generation (G 0), which were heritably transmitted to the following G1 generation, leading to bright red compound eyes and ocelli. Mutations of N. lugens white (Nl-w ) generated a high mutant rate of up to 27.3%, resulting in mosaic eyes consisting of white and lightly pigmented ommatidia in both G 0 and G1 individuals. The specificity of CRISPR/Cas9-mediated mutagenesis was further bolstered by PCR and RNA interference-based knockdown analysis. These results show that CRISPR/Cas9-mediated gene editing is achievable in a hemipteran insect, offering a valuable tool for the study of functional genomics and pest management in this planthopper species. |
Databáze: | OpenAIRE |
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