Direct parental CRISPR gene editing in the predatory bug Orius strigicollis, a biocontrol agent against small arthropods.
Pest Manag Sci
; 80(10): 5465-5472, 2024 Oct.
Article
in En
| MEDLINE
| ID: mdl-38934844
ABSTRACT
BACKGROUND:
The predatory flower bug Orius strigicollis serves as a valuable biocontrol agent against small arthropods; however, its effectiveness can vary, especially when population establishment fails due to low prey/pest densities. A promising approach to improve the efficacy of O. strigicollis as a biocontrol agent is through gene editing. However, as females lay their eggs in plant tissue, the conventional embryo injection approach is challenging in this species.RESULTS:
In this study, we aimed to develop an efficient and practical gene editing technique for O. strigicollis using direct parental CRISPR (DIPA-CRISPR). Female bugs at various postemergence stages received Cas9 ribonucleoprotein injections, with subsequent genotyping of their offspring (G0) using PCR and a heteroduplex mobility assay. We targeted the kynurenine 3-monooxygenase gene (cinnabar), pivotal for insect ommochrome pigment biosynthesis. Through experimental optimization, we achieved a peak gene editing efficiency of 52%, i.e., 52% of G0 progeny carried gene-edited alleles when injecting 1 day postemergence. Notably, some gene-edited G0 adults exhibited a red-eye mosaic phenotype, in contrast to the black-eyed wild type. Crossing experiments confirmed the heritability of the introduced mutations in the subsequent generation (G1), enabling the establishment of a cinnabar-knockout line with bright red eyes.CONCLUSION:
We demonstrate that our DIPA-CRISPR gene editing method tailored for O. strigicollis is efficient and practical. Our findings highlight the potency of DIPA-CRISPR as a tool for O. strigicollis genetic engineering and suggest broader applications for enhancing other biocontrol agents. © 2024 Society of Chemical Industry.Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Pest Control, Biological
/
Heteroptera
/
CRISPR-Cas Systems
/
Gene Editing
Limits:
Animals
Language:
En
Journal:
Pest Manag Sci
Year:
2024
Document type:
Article