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Female-specific insect lethality engineered using alternative splicing

Nature Biotechnology volume 25, pages 353–357 (2007)Cite this article

Abstract

The Sterile Insect Technique is a species-specific and environmentally friendly method of pest control involving mass release of sterilized insects that reduce the wild population through infertile matings1,2,3,4,5. Insects carrying a female-specific autocidal genetic system offer an attractive alternative to conventional sterilization methods6,7 while also eliminating females from the release population7,8,9,10. We exploited sex-specific alternative splicing in insects to engineer female-specific autocidal genetic systems in the Mediterranean fruit fly, Ceratitis capitata. These rely on the insertion of cassette exons from the C. capitata transformer gene into a heterologous tetracycline-repressible transactivator such that the transactivator transcript is disrupted in male splice variants but not in the female-specific one. As the key components of these systems function across a broad phylogenetic range, this strategy addresses the paucity of sex-specific expression systems (e.g., early-acting, female-specific promoters) in insects other than Drosophila melanogaster. The approach may have wide applicability for regulating gene expression in other organisms, particularly for combinatorial control with appropriate promoters.

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Figure 1: Sex-specific expression by alternative splicing of Cctra.
Figure 2: Sex-specific lethal constructs.
Figure 3: Repressible female-specific lethality in transgenic Medflies.
Figure 4: Conservation of function of a Cctra-based construct.

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Acknowledgements

We thank Pedro Rendón (USDA-APHIS-PPQ-CPHST) for the Toliman wild-type strain, and Trudi Mackay for the SamI236 strains; Sinead O'Connell and Veronica Rodrigues for help with D. melanogaster; Karen Clifton and Lynn Richardson for technical assistance and Sinead O'Connell and Derric Nimmo for critical review of the manuscript. This work was funded in part by the UK Biotechnology and Biological Sciences Research Council.

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Author notes

  1. Guoliang Fu and Kirsty C Condon: These authors contributed equally to this work.

Authors and Affiliations

  1. Oxitec Limited, 71 Milton Park, Oxford, OX14 4RX, UK

    Guoliang Fu, Kirsty C Condon, Li Jin, George C Condon, Neil I Morrison, Tarig H Dafa'alla & Luke Alphey

  2. Department of Zoology, University of Oxford, South Parks Road, Oxford, OX1 3PS, UK

    Kirsty C Condon, Matthew J Epton, Peng Gong, Neil I Morrison & Luke Alphey

Authors
  1. Guoliang Fu
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Contributions

G.F. and K.C.C. contributed equally to this work. G.F. and L.J. made the DNA constructs and assisted with the molecular analysis of transgenics. K.C.C. and G.C.C. created the transgenic medfly lines described and conducted most of the phenotypic and molecular analysis. M.J.E. and P.G. performed preliminary experiments in medflies. N.I.M. performed the D. melanogaster experiments. T.H.D. provided advice. L.A. conceived and supervised the project and wrote the paper. All authors discussed the results and commented on the manuscript.

Corresponding author

Correspondence to Luke Alphey.

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Competing interests

Those authors affiliated with Oxitec Ltd. are or were employees or collaborative students of this company, which therefore provided salary and other support for the research program. Also, all these employees have or have had shares or share options in Oxitec Ltd. Both Oxitec Ltd. and Oxford University have one or more patents or patent applications related to the subject of this paper.

Supplementary information

Supplementary Fig. 1

Splicing of Cetra intron in Drosophila melanogaster and Ceratitis capitata. (PDF 203 kb)

Supplementary Fig. 2

Sequence of plasmid pLA3077. (PDF 130 kb)

Supplementary Fig. 3

Sequence of plasmid pLA3097. (PDF 132 kb)

Supplementary Table 1

Repressible female-specific lethality in heterozygous transgenic Medfly. (PDF 127 kb)

Supplementary Table 2

Repressible female-specific lethality in homozygous transgenic Medfly. (PDF 106 kb)

Supplementary Table 3

Repressible female-specific lethality in heterozygous transgenic Drosophila. (PDF 120 kb)

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Fu, G., Condon, K., Epton, M. et al. Female-specific insect lethality engineered using alternative splicing. Nat Biotechnol 25, 353–357 (2007). https://doi.org/10.1038/nbt1283

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