ORIGINAL RESEARCH article
Front. Syst. Biol.
Sec. Integrative Systems Microbiology
This article is part of the Research TopicSystems Biology in Biomedical Innovations and HealthcareView all articles
A Synthetic Biology Toolkit for rational design of genetic circuits in Acinetobacter baumannii
Provisionally accepted
Sara Letrari1
Lisa Faccincani1Ilgin Ertan1Tommaso Varaschin1Francesca Galiazzo1Marco Costanzo2Giorgia D'Angelo3Valentina Del Giudice4Luca Guarnieri5Alex Martini3
Asia Picchi3Chiara Ravazzolo3Niccolò Venturini Degli Esposti5Chiara Zanin3
Livio Trainotti3
Cristiano De Pitta3
Del Vecchio Claudia6
Ignazio Castagliuolo6
Massimo Bellato4,7*- 1Department of Molecular Medicine, University of Padua, Padua, Italy
- 2Department of Medicine, Università degli Studi di Padova, Padova, Italy
- 3Department of Biology, Universita degli Studi di Padova, Padua, Italy
- 4Department of Molecular Medicine, Università degli Studi di Padova, Padova, Italy
- 5Department of Information Engineering, Università degli Studi di Padova, Padova, Italy
- 6Department of Molecular Medicine, Universita degli Studi di Padova, Padua, Italy
- 7Department of Information Engineering, Universita degli Studi di Padova, Padua, Italy
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Antimicrobial resistance (AMR) poses a severe global health threat, with Acinetobacter baumannii among the critical AMR priorities highlighted by the WHO. This Gram-negative pathogen exhibits intrinsic resistance traits, exceptional environmental persistence, and high genomic plasticity harboring resistance islands. To combat AMR through synthetic biology, this work characterizes a library of BioBrick parts to be adopted in A. baumannii engineering, and develops a modular CRISPR interference (CRISPRi) platform. Key components were characterized, including two plasmid vectors, a library of inducible and constitutive promoters, and a CRISPRi-mediated repression system; for the latter, a testbed for biofilm-related genes implicated in the downregulation of antibiotic resistance is also provided. By enabling tunable transcriptional control through the characterized promoters and ensuring the ability to downregulate gene expression via CRISPRi, this synthetic biology toolkit lays the foundation for rational design of genetic circuits to study and counteract AMR in A. baumannii. The modular platform here characterized provides a valuable resource for the iGEM community to advance functional genomics approaches against this alarming global health challenge.
Keywords: Acinetobacter baumannii, Genetic circuit, BioBrick, CRISPRi, Gene Expression, Synthetic & Systems Biology
Received: 18 Jul 2025; Accepted: 05 Nov 2025.
Copyright: © 2025 Letrari, Faccincani, Ertan, Varaschin, Galiazzo, Costanzo, D'Angelo, Del Giudice, Guarnieri, Martini, Picchi, Ravazzolo, Venturini Degli Esposti, Zanin, Trainotti, De Pitta, Claudia, Castagliuolo and Bellato. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Massimo Bellato, massimo.bellato@unipd.it
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