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

Schetelig, Marc F.; Schwirz, Jonas; Yan, Ying

doi:10.1038/s41598-021-91938-1

Cited by 16 publications

(25 citation statements)

References 55 publications

(83 reference statements)

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“…To investigate how SSIMS compares to other genetic biocontrol technologies, we developed an agent-based model for Spotted Wing Drosophila (SWD), an agricultural pest that is the target of genetic biocontrol research and development projects ( Asplen et al, 2015 ; Schetelig et al, 2021 ). Details of the model are described in Supplementary Note 6.…”

Section: Resultsmentioning

confidence: 99%

Genetically engineered insects with sex-selection and genetic incompatibility enable population suppression

Upadhyay

Feltman

Sychla

et al. 2022

eLife

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Engineered Genetic Incompatibility (EGI) is a method to create species-like barriers to sexual reproduction. It has applications in pest control that mimic Sterile Insect Technique when only EGI males are released. This can be facilitated by introducing conditional female-lethality to EGI strains to generate a sex-sorting incompatible male system (SSIMS). Here, we demonstrate a proof of concept by combining tetracycline-controlled female lethality constructs with a pyramus-targeting EGI line in the model insect Drosophila melanogaster. We show that both functions (incompatibility and sex-sorting) are robustly maintained in the SSIMS line and that this approach is effective for population suppression in cage experiments. Further we show that SSIMS males remain competitive with wild-type males for reproduction with wild-type females, including at the level of sperm competition.

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Section: Resultsmentioning

confidence: 99%

Genetically engineered insects with sex-selection and genetic incompatibility enable population suppression

Upadhyay

Feltman

Sychla

et al. 2022

eLife

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“…Previously, crossing V229_M4f1 males with WT females on non-Tet diet produced transgenic offspring consisting of only 14.2% females (Schetelig et al, 2021). When Tet-100 was used here, regardless of the F0 sex and age, the female percentages of transgenic adults were in the range of 14.6%-24.5%, which were significantly lower than those of WT in each cross (p < 0.05, paired t-test) (Figure 3A).…”

Section: Tet and Dox Regulated The Expression Of Tetoff Constructs In...mentioning

confidence: 63%

“…V229_pBXLII_attP_PUbAmCyan_Dsnullo-tTA-SV40_TREhs43-Dshid Ala4 -CctraF-SV40 (Schetelig et al, 2021) at the Bsu36I and MluI restriction sites to generate V359_pBXLII_attP_PUbAmCyan_Dsnullo-tTA-SV40_ TREhs43-DsRed-NLS-DrosCV-2A-EGFP-CAAX-SV40, V360_pBXLII_ attP_PUbAmCyan_Dsnullo-tTA-SV40_TREhs43-DsRed-NLS-TaV-2A-EGFP-CAAX-SV40, V361_pBXLII_attP_PUbAmCyan_Dsnullo-tTA-SV40_TREhs43-EGFP-NLS-DrosCV-2A-DsRed-CAAX-SV40, and V362_ pBXLII_attP_PUbAmCyan_Dsnullo-tTA-SV40_TREhs43-EGFP-NLS-TaV-2A-DsRed-CAAX-SV40, respectively.…”

Section: Insect Rearingmentioning

confidence: 99%

“…The latter is further subjected to the positional effects of the transgene (Schetelig et al, 2016;Concha et al, 2020;Yan et al, 2020c). It is possible that the low female lethality that we previously observed in some V229 lines was due to the low or moderate activity of the pro-apoptotic gene that was further suppressed by parental Tet (Schetelig et al, 2021). Therefore, we selected line V229_M8f2 that showed the lowest female lethality (32.7%) among seven V229 lines in the earlier study for a transgenerational longevity test.…”

Section: Figurementioning

confidence: 99%

“…The Tet-off construct V229 that used to generate these FK strains contains an embryonic promoter from Drosophila suzukii nullo gene to regulate the tTA (driver cassette) and a sex-specifically spliced pro-apoptotic gene hid Ala4 (effector cassette) to eliminate female flies. While the most efficient FK strain eliminated all-female offspring at the embryonic stage if Tet was only fed to the larvae in the parental generation, some other strains killed the majority of females at late stages ( Schetelig et al, 2021 ). It was speculated that Tet could be maternally inherited and switch off the embryonic lethality ( Horn and Wimmer, 2003 ; Schetelig et al, 2009 ; Schetelig and Handler, 2012a ; Yan et al, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

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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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Radioactivity and GMO‐Free Sterile Insect Technology for the Sustainable Control of the Invasive Pest Drosophila suzukii

Abdelhafiz,

Gerth,

Claussen

et al. 2024

Advanced Biology

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Drosophila suzukii (D. suzukii), commonly known as the spotted wing drosophila, is a highly invasive crop pest that is difficult to control using chemical insecticides. To address the urgent need for alternative and more sustainable control strategies, the sterile insect technique (SIT) is improved, which involves the release of sterilized male insects to mate with fertile conspecifics, thereby reducing the size of the pest population in the subsequent generation. The three critical aspects that influence the success of SIT programs in D. suzukii are addressed. First, an accurate and nondestructive method is established to determine the sex of individual insects based on the differential weight of male and female pupae. Second, conditions for X‐ray sterilization are systematically tested and an optimal dose (90 kV/40 Gy) is identified that ensures the efficient production of sterile D. suzukii for release. Finally, the inherent thermosensitivity of D. suzukii males is exploited to develop a temperature‐based sterilization technique, offering an alternative or additional SIT method for this pest. These advances will contribute to the development of a comprehensive and effective strategy for the management of D. suzukii populations, reducing their impact on agriculture and helping to safeguard crop yields.

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

Cited by 16 publications

References 55 publications

Genetically engineered insects with sex-selection and genetic incompatibility enable population suppression

Genetically engineered insects with sex-selection and genetic incompatibility enable population suppression

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

Radioactivity and GMO‐Free Sterile Insect Technology for the Sustainable Control of the Invasive Pest Drosophila suzukii

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