Selective gene expression in hydrogel suspended escherichia coli through optogenetics by spatiotemporally controlled light for the purposes of additive manufacturing
File(s)
Author(s)
Lampret, Borut
Type
Thesis
Abstract
Advances in the ability to manipulate genetic material have enabled the utilization of microbes in production of both biological and synthetic materials. However, the vast majority of these materials are not suitable for additive manufacturing. While there have been significant advances in the field of bioprinting, significant effort is required to develop the so called bioinks, which support both the printing process and the encapsulated cells’ survivability. To combat these challenges, the work in this thesis investigates a novel method of additive manufacturing, which can employ microbial cell factories suspended in hydrogel and optogenetics to selectively deposit the desired biological or synthetic material in a three-dimensional volume.
Genetically engineered Escherichia coli that produces CcaSR, a synthetic light response system, was used in order to establish spatially selective gene expression. A custom designed system was built to house the hydrogel suspended E. coli and to provide and shape the green and red light used to interact with the cells. A series of experiments were carried out to confirm localized gene expression within the hydrogel build volume and to test and confirm the behaviour of the CcaSR system. To enable the process of additive manufacturing, a series of spatiotemporal illumination procedures were developed and their behaviour investigated. A select illumination procedure was then employed in inducing gene expression in a nontrivial three-dimensional object confirming the viability of the proposed method for the purposes of additive manufacturing.
Genetically engineered Escherichia coli that produces CcaSR, a synthetic light response system, was used in order to establish spatially selective gene expression. A custom designed system was built to house the hydrogel suspended E. coli and to provide and shape the green and red light used to interact with the cells. A series of experiments were carried out to confirm localized gene expression within the hydrogel build volume and to test and confirm the behaviour of the CcaSR system. To enable the process of additive manufacturing, a series of spatiotemporal illumination procedures were developed and their behaviour investigated. A select illumination procedure was then employed in inducing gene expression in a nontrivial three-dimensional object confirming the viability of the proposed method for the purposes of additive manufacturing.
Version
Open Access
Date Issued
2022-12
Date Awarded
2023-03
Copyright Statement
Creative Commons Attribution NonCommercial NoDerivatives Licence
Advisor
Myant, Connor
Stan, Guy-Bart
Publisher Department
Department of Chemistry; Dyson School of Design Engineering
Publisher Institution
Imperial College London
Qualification Level
Doctoral
Qualification Name
Doctor of Philosophy (PhD)