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Assessing the Orthogonality of Genetic Circuits
Abu-Ageel, Beteel
Abu-Ageel, Beteel
Abstract
Orthogonality is one of the most fundamental principles of synthetic biology. Designing circuits that limit unintended interactions between a circuit and the genetic background of its host is essential for ensuring host health, circuit functionality, and overall safety of the engineered organism (Borkowski et al., 2016). However, a standardized method of orthogonality assessment between a genetic circuit and its host does not currently exist in the field. In addition, some of the most effective methods for assessing circuit-host orthogonality, such as RNA sequencing, are not widely accessible due to the technical expertise required and the high financial cost. This thesis aims to provide a review of the scientific literature in which orthogonality of a genetic circuit has been implicitly or explicitly assessed in prokaryotic organisms, and also aims to highlight some of the methods and tools that have been developed for increased circuit orthogonalization (Costello & Badran, 2021; Liu et al., 2018b). In addition to this review, this thesis summarizes the circuit design finalization, construction, and testing that Beteel Abu-Ageel completed as a member of the 2021 William and Mary iGEM team. Specifically, this includes her work with sensors designed to quantify burden caused by a circuit on both the transcriptional and translational levels and testing of a curli fiber production circuit which served as a test circuit for the burden sensor experiments.
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2022-12-01
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Embargoed through December 12, 2025
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Biology