A Synthetic Biology Approach to Transgene Expression in Insects

• Published in: ACS synthetic biology 2024-09, Vol.13 (9), p.3041-3045
• Main Authors: Leftwich, Philip T., Purcell, Jessica C., Anderson, Michelle A. E., Fragkoudis, Rennos, Basu, Sanjay, Lycett, Gareth, Alphey, Luke
• Format: Article
• Language: English
• Subjects: 3' Untranslated Regions - genetics; Animals; Animals, Genetically Modified; Cell Line; Culicidae - genetics; Gene Expression - genetics; Gene Expression Regulation - genetics; Genetic Engineering - methods; Insecta - genetics; Peptide Chain Initiation, Translational - genetics; Promoter Regions, Genetic - genetics; RNA Polymerase II - genetics; RNA Polymerase II - metabolism; Synthetic Biology - methods; Technical Note; Transgenes
• ISSN: 2161-5063; 2161-5063
• DOI: 10.1021/acssynbio.4c00250

The ability to control gene expression is pivotal in genetic engineering and synthetic biology. However, in most nonmodel and pest insect species, empirical evidence for predictable modulation of gene expression levels is lacking. This knowledge gap is critical for genetic control systems, particularly in mosquitoes, where transgenic methods offer novel routes for pest control. Commonly, the choice of RNA polymerase II promoter (Pol II) is the primary method for controlling gene expression, but the options are limited. To address this, we developed a systematic approach to characterize modifications in translation initiation sequences (TIS) and 3′ untranslated regions (UTR) of transgenes, enabling the creation of a toolbox for gene expression modulation in mosquitoes and potentially other insects. The approach demonstrated highly predictable gene expression changes across various cell lines and 5′ regulatory sequences, representing a significant advancement in mosquito synthetic biology gene expression tools.