A synthetic biology approach to transgene expression in insects

Leftwich, Philip T. ORCID: https://orcid.org/0000-0001-9500-6592, Purcell, Jessica C., Anderson, Michelle A. E., Fragkoudis, Rennos, Basu, Sanjay, Lycett, Gareth and Alphey, Luke (2024) A synthetic biology approach to transgene expression in insects. ACS Synthetic Biology, 13 (9). 3041–3045. ISSN 2161-5063

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Abstract

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.

Item Type:	Article
Additional Information:	For the purpose of open access, the author has applied a CC BY public copyright licence to any Author Accepted Manuscript version arising from this submission. The findings and conclusions contained within are those of the authors and do not necessarily reflect positions or policies of the Bill & Melinda Gates Foundation. The views, opinions and/or findings expressed are those of the authors and should not be interpreted as representing the official views or policies of the Department of Defense or the U.S. Government. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Data Availability Statement: Scripts and raw data can be found at Github (https://github.com/Philip-Leftwich/Pol2-promoters). Complete information on analyses can be found in Supporting Information. Funding information: This research was funded in part by research grants from the Wellcome Trust [110117/Z/15/Z and 226721/Z/22/Z] and the Bill & Melinda Gates Foundation [INV-008549] to L.A. and by the Defense Advanced Research Projects Agency (DARPA) Safe Genes Program [N66001-17-2-4054] to Kevin Esvelt at MIT. J.P. was supported by a studentship from The Pirbright Institute (BBS/E/I/00001985). L.A. was also supported through strategic funding from the UK Biotechnology and Biological Sciences Research Council (BBSRC) to The Pirbright Institute (BBS/E/I/00007033, BBS/E/I/00007038 and BBS/E/I/00007039).
Faculty \ School:	Faculty of Science > School of Biological Sciences
UEA Research Groups:	Faculty of Science > Research Groups > Biosciences Teaching and Education Research
Related URLs:	https://pubs.acs.org/doi/10.1021/acssynb...
Depositing User:	LivePure Connector
Date Deposited:	29 Aug 2024 09:30
Last Modified:	14 Oct 2024 19:30
URI:	https://ueaeprints.uea.ac.uk/id/eprint/96382
DOI:	10.1021/acssynbio.4c00250

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