Identification of genes for engineering the male germline of Aedes aegypti and Ceratitis capitata

Background 27 Synthetic biology approaches are promising new strategies for control of pest insects 28 that transmit disease and cause agricultural damage. These strategies require 29 characterised modular components that can direct appropriate expression of effector 30 sequences, with components conserved across species being particularly useful. 31 The goal of this study was to identify genes from which new potential components 32 could be derived for manipulation of the male germline in two major pest species, the 33 mosquito Aedes aegypti and the tephritid fruit fly Ceratitis capitata. Results 36 Using RNA-seq data from staged testis samples, we identified several candidate 37 genes with testis-specific expression and suitable expression timing for use of their 38 regulatory regions in synthetic control constructs. We also developed a novel 39 computational pipeline to identify candidate genes with testis-specific splicing from 40 this data; use of alternative splicing is another method for restricting expression in 41 synthetic systems. Some of the genes identified display testis-specific expression or 42 splicing that is conserved across species; these are particularly promising candidates 43 for construct development. Conclusions 46 In this study we have identified a set of genes with testis-specific expression or 47 splicing. In addition to their interest from a basic biology perspective, these findings 48 provide a basis from which to develop synthetic systems to control important pest 49 insects via manipulation of the male germline