First Anopheles arabiensis germline transformation: Toward the development of a transgenic genetic sexing strain
First Anopheles arabiensis germline transformation: Toward the development of a transgenic genetic sexing strain
Tags: Anopheles, Genetically modified mosquitoes, Malaria, Sterile insect technique (SIT)H. C. Bossin, J. Thailayil, F. Catteruccia, J. P. Benton, A. Crisanti, M. Q. Benedict, B. G. Knols and A. S. Robinson, American Journal of Tropical Medicine and Hygiene, 75:66-66. 2006.
The ability to genetically engineer mosquitoes is likely to have major implications for the development and implementation of genetic control systems against mosquito disease vectors such as the Sterile Insect Technique (SIT). In particular, genetically transformed mosquito strains can be created for genetic marking and sexing, two key factors known to influence the effectiveness of SIT programmes. In addition, the removal of biting females before releasing sterile males in the field will be of critical importance as they contribute to disease transmission and reduce the efficiency of the release campaign. Parallel to the creation of a conventional genetic sexing strain (Y-translocation of a resistance marker), our group is undertaking a transgenic approach to the development of an A. arabiensis genetic sexing strain (GSS). The sex separation strategy under investigation relies on the sex-specific properties of the A. gambiae B2tubulin gene regulatory regions. It is hoped this approach will achieve the high sex separation efficiency (above 99%) and strain stability required for safe and efficient male-only SIT releases. We report here the successful development of transgenic A. arabiensis lines using the pPB[DsRed]B2EGFP construct. Wild-type A. arabiensis embryos were injected with a mixture of pPB[DsRed]B2EGFP and helper plasmid phsppBac (700 and 300 ng/µI respectively) following an appropriate protocol. Injections generated several transgenic sexing lines. The effectiveness of the transgenic-based sex-separation procedure, the stability of transgenic mosquito GSS under various (mass-)rearing regimes, as well as the viability and reproductive competitiveness of transgenic sterile males are being assessed.