D3367: Fitness impact of transgene elements in Cas9-expressing transgenic Anopheles stephensi lines

Transgenic mosquito approaches to disease control are designed to equip the mosquito genome with new synthetic gene combinations that cause either a local mosquito population crash (population repression) or blocking of human pathogen transmission within the mosquito (population modification). At the laboratory end of these efforts, genetic engineers design one or several transgenes, the set of DNA code assembled of gene parts from a number of organisms including other eukaryotes, bacteria, viruses and phages. Fitness effects of transgene sequences have the potential to impact downstream applications but can be challenging to measure. We have used assayed life table parameters in a set of transgenic Anopheles stephensi with similar, but variable, synthetic transgene constructs at different insertion locations to test fitness effects of different parts of commonly used transgene elements including fluorescent markers and the gene-editing bacterial endonuclease Cas9. Such data will support more accurate predictions for downstream deployment of new transgenic technologies for applications in mosquito-borne disease.

Key words – Mosquito-borne disease, Fitness impact, insertion locations, gene-editing bacterial endonuclease Cas9.