Conditional Expression Systems for Drosophila suzukii Pest Control

Jaffri, Syeda A.; Yan, Ying; Scott, Maxwell J.; Schetelig, Marc F.

doi:10.1007/978-3-030-62692-1_10

Cited by 5 publications

(2 citation statements)

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“…A tissue-specific CRISPR using the UAS/Gal4 system has been developed in D. melanogaster, which restricts mutagenesis to certain tissues or cells, thus rescuing individuals that carry lethal mutations [54,55]. Alternatively, other conditional expression systems such as those regulated by antibiotics, non-antibiotic molecules, or external stimuli could be considered for the control of Cas9 expression [56]. Our work here verified that the Cas9 expression in the Dmhsp70-Cas9 line is heat-inducible, and its basal expression level is associated with the genomic position of the transgene.…”

Section: Discussionmentioning

confidence: 99%

Highly Efficient Temperature Inducible CRISPR-Cas9 Gene Targeting in Drosophila suzukii

Yan

Kobayashi

Huang

et al. 2021

IJMS

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The spotted-wing Drosophila (Drosophila suzukii Matsumura) is native to eastern Asia, but has become a global threat to fruit production. In recent years, CRISPR/Cas9 targeting was established in this species allowing for functional genomic and genetic control studies. Here, we report the generation and characterization of Cas9-expressing strains of D. suzukii. Five independent transgenic lines were generated using a piggyBac construct containing the EGFP fluorescent marker gene and the Cas9 gene under the control of the D. melanogaster heat shock protein 70 promoter and 3’UTR. Heat-shock (HS) treated embryos were analyzed by reverse transcriptase PCR, revealing strong heat inducibility of the transgenic Cas9 expression. By injecting gRNA targeting EGFP into one selected line, 50.0% of G0 flies showed mosaic loss-of-fluorescence phenotype, and 45.5% of G0 flies produced G1 mutants without HS. Such somatic and germline mutagenesis rates were increased to 95.4% and 85.7%, respectively, by applying a HS. Parental flies receiving HS resulted in high inheritance of the mutation (92%) in their progeny. Additionally, targeting the endogenous gene yellow led to the lack of pigmentation and male lethality. We discuss the potential use of these efficient and temperature-dependent Cas9-expressing strains for the genetic studies in D. suzukii.

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Section: Discussionmentioning

confidence: 99%

Highly Efficient Temperature Inducible CRISPR-Cas9 Gene Targeting in Drosophila suzukii

Yan

Kobayashi

Huang

et al. 2021

IJMS

Self Cite

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“…It is therefore key to find control methods focusing mostly on females (e.g. Jaffri et al, 2020), which seem to be the most problematic since they can disperse over longer distances.…”

Section: Implications For Management In Raspberry Orchardsmentioning

confidence: 99%