Using Moderate Transgene Expression to Improve the Genetic Sexing System of the Australian Sheep Blow Fly Lucilia cuprina

Yan, Ying; Williamson, Megan E; Scott, Maxwell J.

doi:10.3390/insects11110797

Cited by 5 publications

(3 citation statements)

References 36 publications

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“…Therefore, the driver and effector cassettes from the V229 vector can be arranged into two constructs which can be used to generate independent transgenic lines. By evaluating the activities of tTA and pro-apoptotic gene in these driver and effector lines, respectively, it should be possible to develop an FK strain without the leaky lethality by choosing driver and effector lines with moderate transgene activities 43 .…”

Section: Discussionmentioning

confidence: 99%

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: 99%

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

Schetelig

Schwirz

Yan

2021

Sci Rep

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“…It was recognized that the parental antibiotics might suppress the lethality correlated with the pro-apoptotic gene’s activity or expression level ( Horn and Wimmer, 2003 ; Schetelig et al, 2009 ; Yan and Scott, 2015 ; Yan et al, 2017 ). 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 ).…”

Section: Resultsmentioning

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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“…The SIT has also been proposed as a tool to control populations of another major livestock pest species, the Australian sheep blow fly Lucilia cuprina . Yan and colleagues (2020) [ 25 ] report on the development of a transgenic embryonic sexing system (TESS) that results in the elimination of females at the embryonic stage in the absence of tetracycline from the diet, which could potentially be used for male-only releases if regulatory approval is granted.…”

Section: Livestock Pestsmentioning

confidence: 99%

Sterile Insect Technique (SIT) and Its Applications

Bourtzis¹,

Vreysen²

2021

Insects

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Although most insect species have a beneficial role in the ecosystems, some of them represent major plant pests and disease vectors for livestock and humans. During the last six–seven decades, the sterile insect technique (SIT) has been used as part of area-wide integrated pest management strategies to suppress, contain, locally eradicate or prevent the (re)invasion of insect pest populations and disease vectors worldwide. This Special Issue on “Sterile insect technique (SIT) and its applications”, which consists of 27 manuscripts (7 reviews and 20 original research articles), provides an update on the research and development efforts in this area. The manuscripts report on all the different components of the SIT package including mass-rearing, development of genetic sexing strains, irradiation, quality control as well as field trials.

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Using Moderate Transgene Expression to Improve the Genetic Sexing System of the Australian Sheep Blow Fly Lucilia cuprina

Cited by 5 publications

References 36 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

Sterile Insect Technique (SIT) and Its Applications

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