Engineering Novel Bandpass Tools for Cascading Gene Expression
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Onajobi, Glory Jesupelumi
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Abstract
Genetic circuits are synthetic biological systems engineered to perform specific functions within living organisms. These circuits play a crucial role in biomanufacturing, optimizing metabolic pathways, and enhancing product yield. With the continuous expansion of genetic circuit engineering, there is an increasing demand for introducing and applying advanced circuits that leverage modularity to improve overall efficiency. This thesis aims to construct a novel compound logical operation called the Cascade, designed for system optimization and precise, conditional control. The Cascade implements the architecture of a triple sequential logic operation capable of processing four-signal inputs, regulating three different genes, and transitioning gene expression to facilitate their OFF-ON-OFF states based on the sequence of inputs introduced. This intricate logical operation represents the fusion of three NIMPLY logic gates for cascaded gene expression, where each repressor-anti-repressor pair is directed to regulate the expression of a single gene. To achieve this fusion, a series of foundational logic operations were built and tested. These included single-input single-output (SISO) logic operations and multiple-input single-output (MISO) logic operations, providing valuable insights into the efficiency and functionality of this fusion. The design and implementation of the Cascade qualitatively regulated three different segments in a sequential pathway, altered regulatory phenotype and function without small molecule removal, and enabled precise and conditional control of the system. These aspects of the sequential logic operation make it a valuable tool for optimizing flux for metabolic pathways and similar processes.
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2024-07-08
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Thesis