Functional characterization of the Drosophila suzukii pro-apoptotic genes reaper, head involution defective and grim

Jaffri, Syeda A.; Yan, Ying; Schwirz, Jonas; Schetelig, Marc F.

doi:10.1007/s10495-020-01640-2

Cited by 4 publications

(5 citation statements)

References 60 publications

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“…We therefore selected the Dsnullo and Dssry-α promoters to control tTA expression in the driver cassette of our AIO constructs. Similarly, Dsgrim and Dshid were shown to be more potent pro-apoptotic genes than Dsrpr in S2 cell death assays 29 . We therefore used Dsgrim and Dshid Ala4 , the latter predicted to be more lethal than the endogenous version 18 , 21 , in our effector cassette.…”

Section: Discussionmentioning

confidence: 94%

“…Each reaction was heated to 98 °C for 3 min followed by 40 cycles of 98 °C for 10 s and 60 °C for 30 s. The primers are listed in Table S2 . Three biological and three technical replicates were carried out for each reaction, and results were normalized to the TATA binding protein ( TBP ) reference gene using the 2 -∆∆Ct method as previously described 28 , 29 .…”

Section: Methodsmentioning

confidence: 99%

“…The female-specific intron from a sex determination gene such as transformer ( tra ) is typically placed within a pro-apoptotic gene (effector cassette) to ensure that only females are killed 18 , 19 , 22 , 23 . We have previously reported the isolation and analysis of D. suzukii cellularization gene promoters, pro-apoptotic genes, and the nuclear localization signal (NLS) of the tra gene from the standard USA laboratory strain 27 – 29 . In these studies, we confirmed the embryonic expression of the cellularization genes nullo, serendipity-α ( sry-α ), bottleneck ( bnk ), and slow-as-molasses ( slam ) and the functionality of the pro-apoptotic genes head involution defective ( hid ), grim, and reaper ( rpr ) .…”

Section: Introductionmentioning

confidence: 99%

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A transgenic female killing system for the genetic control of Drosophila suzukii

Schetelig

Schwirz

Yan

2021

Sci Rep

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The spotted wing Drosophila (Drosophila suzukii) is an invasive pest of soft-skinned fruit crops. It is rapidly transmitted in Europe and North America, causing widespread agricultural losses. Genetic control strategies such as the sterile insect technique (SIT) have been proposed as environment-friendly and species-restricted approaches for this pest. However, females are inefficient agents in SIT programs. Here we report a conditional female-killing (FK) strategy based on the tetracycline-off system. We assembled sixteen genetic constructs for testing in vitro and in vivo. Twenty-four independent transgenic strains of D. suzukii were generated and tested for female-specific lethality. The strongest FK effect in the absence of tetracycline was achieved by the construct containing D. suzukii nullo promoter for early gene expression, D. suzukii pro-apoptotic gene hidAla4 for lethality, and the transformer gene intron from the Mediterranean fruit fly Ceratitis capitata for female-specific splicing. One strain carrying this construct eliminated 100% of the female offspring during embryogenesis and produced only males. However, homozygous females from these FK strains were not viable on a tetracycline-supplemented diet, possibly due to the basal expression of hidAla4. Potential improvements to the gene constructs and the use of such FK strains in an SIT program are discussed.

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Section: Discussionmentioning

confidence: 94%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A transgenic female killing system for the genetic control of Drosophila suzukii

Schetelig

Schwirz

Yan

2021

Sci Rep

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“…Both DrosCV-2A and TaV-2A have been previously used to co-express two different pro-apoptotic genes including reaper , head involution defective ( hid ), and grim . S2 cell death assays showed that the lethality can be significantly enhanced by such binary expression compared to the single expression of a pro-apoptotic gene ( Jaffri et al, 2020 ). Here, our observation suggested that TaV-2A may be preferred over DrosCV-2A for multiple expression of different lethal genes using the Tet-off system.…”

Section: Resultsmentioning

confidence: 99%

“…However, it may also speed up the generation of the second-site resistance mutations which caused the genetic breakdown of RIDL lethality in D. melanogaster ( Zhao et al, 2020 ). Therefore, incorporating two or more lethal effectors/systems into the insect strains intended for use may help ensure the complete penetrance of lethality and reduce the development of resistance ( Jaffri et al, 2020 ; Yan and Scott, 2020 ; Li et al, 2021 ; Upadhyay et al, 2022 ). For any genetic control strategies employing the Tet-off system, excessive use of Tet or its derivatives could reduce the strain fitness or lead to survivors, which may cause undesired damage or reduce the control efficiency.…”

Section: Discussionmentioning

confidence: 99%

Effects of antibiotics on the in vitro expression of tetracycline-off constructs and the performance of Drosophila suzukii female-killing strains

Yan

Hosseini²,

Scheld³

et al. 2023

Front. Bioeng. Biotechnol.

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Genetic control strategies such as the Release of Insects Carrying a Dominant Lethal (RIDL) gene and Transgenic Embryonic Sexing System (TESS) have been demonstrated in the laboratory and/or deployed in the field. These strategies are based on tetracycline-off (Tet-off) systems which are regulated by antibiotics such as Tet and doxycycline (Dox). Here, we generated several Tet-off constructs carrying a reporter gene cassette mediated by a 2A peptide. Different concentrations (0.1, 10, 100, 500, and 1,000 μg/mL) and types (Tet or Dox) of antibiotics were used to evaluate their effects on the expression of the Tet-off constructs in the Drosophila S2 cells. One or both of the two concentrations, 100 and 250 μg/mL, of Tet or Dox were used to check the influence on the performances of a Drosophila suzukii wild-type strain and female-killing (FK) strains employing TESS. Specifically, the Tet-off construct for these FK strains contains a Drosophila suzukii nullo promoter to regulate the tetracycline transactivator gene and a sex-specifically spliced pro-apoptotic gene hidAla4 to eliminate females. The results suggested that the in vitro expression of the Tet-off constructs was controlled by antibiotics in a dose-dependent manner. ELISA experiments were carried out identifying Tet at 34.8 ng/g in adult females that fed on food supplemented with Tet at 100 μg/mL. However, such method did not detect Tet in the eggs produced by antibiotic-treated flies. Additionally, feeding Tet to the parents showed negative impact on the fly development but not the survival in the next generation. Importantly, we demonstrated that under certain antibiotic treatments females could survive in the FK strains with different transgene activities. For the strain V229_M4f1 which showed moderate transgene activity, feeding Dox to fathers or mothers suppressed the female lethality in the next generation and feeding Tet or Dox to mothers generated long-lived female survivors. For the strain V229_M8f2 which showed weak transgene activity, feeding Tet to mothers delayed the female lethality for one generation. Therefore, for genetic control strategies employing the Tet-off system, the parental and transgenerational effects of antibiotics on the engineered lethality and insect fitness must be carefully evaluated for a safe and efficient control program.

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Biotechnology-enhanced genetic controls of the global pest Drosophila suzukii

Yan,

Ahmed,

Wimmer

et al. 2024

Trends in Biotechnology

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Functional characterization of the Drosophila suzukii pro-apoptotic genes reaper, head involution defective and grim

Cited by 4 publications

References 60 publications

A transgenic female killing system for the genetic control of Drosophila suzukii

A transgenic female killing system for the genetic control of Drosophila suzukii

Effects of antibiotics on the in vitro expression of tetracycline-off constructs and the performance of Drosophila suzukii female-killing strains

Biotechnology-enhanced genetic controls of the global pest Drosophila suzukii

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