Tuning gene expression in mammalian cells by transcriptional and post-transcriptional control
File(s)
Author(s)
Zouein, Annalise
Type
Thesis
Abstract
Engineering biology also known as synthetic biology is rapidly advancing, offering numerous genetic tools for the engineering and reprogramming of cells. These tools enhance cell performance, enabling novel functions, with a wide range of potential applications. The future of cell engineering would benefit from strategies that facilitate precise and tuneable modulation of gene expression without permanent genetic alteration or a large resource footprint. In this direction, we explore the use of microRNAs (miRNAs) and Transcription Factor Binding Sites (TFBS) to control gene expression at transcriptional and post-transcriptional levels. Inspired by favourable native characteristics that ensure tight regulation within cells, we develop a highly tuneable, modular and programmable platform for the control of gene expression in mammalian cells.
The developed miRNAs and TF decoy tools show promise in modulating gene expression in mammalian cells. The use of TF decoys, empowered by our in-house DNA assembly method, led to the sequestration of intracellular TFs, subsequently causing changes in gene expression and cellular responses. On the other hand, sRNAs such as miRNAs were used to alter transcriptional and post-transcriptional outputs of genes showing their versatility. Further development may be required to fully harness their potential for gene expression modulation in mammalian cells.
This work contributes to the growing field of biology-inspired cell engineering, providing insights into the precise tuning of gene expression levels in mammalian cells. The development of these tools opens possibilities for studying diseases, cellular reprogramming for cell therapies and the production of relevant recombinant products.
The developed miRNAs and TF decoy tools show promise in modulating gene expression in mammalian cells. The use of TF decoys, empowered by our in-house DNA assembly method, led to the sequestration of intracellular TFs, subsequently causing changes in gene expression and cellular responses. On the other hand, sRNAs such as miRNAs were used to alter transcriptional and post-transcriptional outputs of genes showing their versatility. Further development may be required to fully harness their potential for gene expression modulation in mammalian cells.
This work contributes to the growing field of biology-inspired cell engineering, providing insights into the precise tuning of gene expression levels in mammalian cells. The development of these tools opens possibilities for studying diseases, cellular reprogramming for cell therapies and the production of relevant recombinant products.
Version
Open Access
Date Issued
2024-01-26
Date Awarded
2024-04-01
Copyright Statement
Creative Commons Attribution NonCommercial NoDerivatives Licence
Advisor
Ellis, Tom
Ceroni, Francesca
Sponsor
Engineering and Physical Sciences Research Council
Grant Number
EP/S022856/1
Publisher Department
Bioengineering
Publisher Institution
Imperial College London
Qualification Level
Doctoral
Qualification Name
Doctor of Philosophy (PhD)