Understanding and improving transgene stability and expression in insects for SIT and conditional lethal release programs

Abstract

Genetically transformed insect pests provide significant opportunities to create strains for improved sterile insect technique and new strategies based on conditional lethality. A major concern for programs that rely on the release of transgenic insects is the stability of the transgene, and maintenance of consistent expression of genes of interest within the transgene. Transgene instability would influence the integrity of the transformant strain upon which the effectiveness of the biological control program depends. Loss or intra-genomic transgene movement would result in strain attributes important to the program being lost or diminished, and the mass-release of such insects could significantly exacerbate the insect pest problem. Instability resulting in intra-genomic movement may also be a prelude to inter-genomic transgene movement between species resulting in ecological risks. This is less of a concern for short-term releases, where transgenic insects are not expected to survive in the environment beyond two or three generations. Transgene movement may occur, however, into infectious agents during mass-rearing, and the potential for movement after release is a possibility for programs using many millions of insects. The primary methods of addressing potential transgene instability relate to an understanding of the vector system used for gene transfer, the potential for its mobilization by the same or a related vector system, and methods required to identify transformants and determine if unexpected transgene movement has occurred. Methods also exist for preventing transposon-mediated mobilization, by deleting or rearranging vector sequences required for transposition using recombination systems. Stability of transgene expression is also a critical concern, especially in terms of potential epigenetic interactions with host genomes resulting in gene silencing that have been observed in plants and fungi, and it must be determined if this or related phenomena can occur in insects.