CRISPR/Cas9-mediated genome editing of Frankliniella occidentalis, the western flower thrips, via embryonic microinjection

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Pubblicato in:bioRxiv (Dec 1, 2023)
Autore principale: Han, Jinlong
Altri autori: Klobasa, William, Lucas De Oliveira, Rotenberg, Dorith, Whitfield, Anna E, Lorenzen, Marce D
Pubblicazione:
Cold Spring Harbor Laboratory Press
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001 2895911941
003 UK-CbPIL
022 |a 2692-8205 
024 7 |a 10.1101/2023.11.30.569456  |2 doi 
035 |a 2895911941 
045 0 |b d20231201 
100 1 |a Han, Jinlong 
245 1 |a CRISPR/Cas9-mediated genome editing of Frankliniella occidentalis, the western flower thrips, via embryonic microinjection 
260 |b Cold Spring Harbor Laboratory Press  |c Dec 1, 2023 
513 |a Working Paper 
520 3 |a The western flower thrips, Frankliniella occidentalis, poses a significant challenge in global agriculture as a notorious pest and a vector of economically significant orthotospoviruses. However, the limited availability of genetic tools for F. occidentalis hampers the advancement of functional genomics and the development of innovative pest control strategies. In this study, we present a robust methodology for generating heritable mutations in F. occidentalis using the CRISPR/Cas9 genome editing system. Two eye-color genes, white (Fo-w) and cinnabar (Fo-cn), frequently used to assess Cas9 function in insects were identified in the F. occidentalis genome and targeted for knockout through embryonic microinjection of Cas9 complexed with Fo-w or Fo-cn specific guide RNAs. Homozygous Fo-w and Fo-cn knockout lines were established by crossing mutant females and males. The Fo-w knockout line revealed an age-dependent modification of eye-color phenotype. Specifically, while young larvae exhibit ivory-colored eyes, the color transitions to bright red as they age. Unexpectedly, loss of Fo-w function also altered body color, with Fo-w mutants having a lighter colored body than wild type, suggesting a dual role for Fo-w in thrips. In contrast, individuals from the Fo-cn knockout line consistently displayed bright red eyes throughout all life stages. Molecular analyses validated precise editing of both target genes. This study offers a powerful tool to investigate thrips gene functions and paves the way for the development of genetic technologies for population suppression and/or population replacement as a means of mitigating virus transmission by this vector.Competing Interest StatementThe authors have declared no competing interest. 
653 |a Color 
653 |a Embryos 
653 |a CRISPR 
653 |a Genomes 
653 |a Pest control 
653 |a Microinjection 
653 |a Mutants 
653 |a Population genetics 
653 |a Phenotypes 
653 |a Frankliniella occidentalis 
700 1 |a Klobasa, William 
700 1 |a Lucas De Oliveira 
700 1 |a Rotenberg, Dorith 
700 1 |a Whitfield, Anna E 
700 1 |a Lorenzen, Marce D 
773 0 |t bioRxiv  |g (Dec 1, 2023) 
786 0 |d ProQuest  |t Biological Science Database 
856 4 1 |3 Citation/Abstract  |u https://www.proquest.com/docview/2895911941/abstract/embedded/7BTGNMKEMPT1V9Z2?source=fedsrch 
856 4 0 |3 Full text outside of ProQuest  |u https://www.biorxiv.org/content/10.1101/2023.11.30.569456v1