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Synthetic genetic circuit engineering: principles, advances and prospects
GAO Ge, BIAN Qi, WANG Baojun
Synthetic Biology Journal    2025, 6 (1): 45-64.   DOI: 10.12211/2096-8280.2023-096
Abstract   (1477 HTML169 PDF(pc) (3188KB)(1802)  

Synthetic genetic circuits are engineered gene networks comprised of redesigned genetic parts for interacting to perform customized functions in cells. With the rapid development of synthetic biology, synthetic genetic circuits have shown significant application potentials in many fields such as biomanufacturing, healthcare and environmental monitoring. However, the efforts to scale up genetic circuits are hindered by the limited number of orthogonal parts, the difficulty of functionally composing large-scale circuits, and the poor predictability of circuit behaviors. A longstanding goal of synthetic biology research is to engineer complex synthetic biological circuits, using modular genetic parts, as we do with electronic circuits. Synthetic biologists have developed various genetic toolboxes and functional assembly methods over the past few decades. Here we present an overview of the latest advances, challenges, and future prospects in genetic circuit engineering from four aspects corresponding to the four key engineering principles for circuit design, i.e. orthogonality, standardization, modularity, and automation. Firstly, the design and construction of orthogonal genetic part libraries are discussed in both prokaryotes and eukaryotes at the levels of DNA replication, transcription, and translation, respectively. Standardized characterization methods and the design of modular genetic parts are subsequently summarized. Furthermore, progress in developing modular genetic circuits are presented, providing new concepts and ways for engineering increasingly large and complex circuits. Finally, how to achieve automated design and building of genetic circuits are addressed from the advances in software, hardware and artificial intelligence, respectively, with an aim to replacing the presently time-consuming manual trial-and-error mode with the iterative "design-build-test-learn" cycle for improved efficiency and predictability of circuit design. The integration of these fundamental principles and the latest advances in information technology such as artificial intelligence and lab automation will accelerate the paradigm shift in genetic circuit engineering and synthetic biology research, making it feasible for designing synthetic lives to meet various customized needs.


Fig. 1 Validated orthogonal parts and tools for precise gene expression control in genetic circuit design
(Synthetic genetic parts can regulate various steps of gene expression, including DNA storage and replication[14-20], transcription[21-45], translation[46-47], and post-translational regulation[48-50].)
Extracts from the Article
生物正交反应是指能够在生物体系中进行,且不会与天然生物化学过程相互干扰的一类化学反应[13]。在基因线路设计过程中,为了精准地调控细胞内各组分的活性,应尽量减少元件之间的相互干扰,这就是基因线路中的“正交化”。随着合成生物学的快速发展,科学家们在遗传信息表达及生化代谢的不同过程中成功创建了正交元件(图1和表1)。
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