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Engineering φX174 to create a biocontained phage therapy model

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posted on 2022-10-14, 01:38 authored by Pamela Tsoumbris

Antibiotic resistant bacteria are an increasing concern amongst hospitals worldwide, with many patients presenting infections that are nonresponsive to current treatments. To resolve this problem, many scientists are turning to phage therapy for treatment. Unfortunately, phage therapy harbours drawbacks including fear of gene transfer between different microbes within human microbiomes, and the potential for off-target interactions interrupting other microbial niches within the body. Engineered phage may reduce some of these drawbacks, creating phage with broader host range or the ability to degrade biofilms, but to realise these benefits outside the lab, engineered phage need to show they cannot spread widely in the environment. In this thesis I created a biocontained phage therapy model to attempt to address these concerns. In employing the non-canonical amber start codon (TAG) at the beginning of φX174 capsid genes, I engineered a phage that could only replicate when provided with the mutant initiator tRNACUA. As a result, I was able to provide a method of biocontainment which maintained phage stability, maintained its ability to adsorb to host cells, and prevented it from replicating, although host-killing efficiency was reduced from wild-type phage. This study provides a platform technology for the effective containment of bacteriophages and takes the first step towards enabling phage therapy to become an accessible treatment option.

History

Table of Contents

Chapter 1: Introduction -- Chapter 2: Methods and Materials -- Chapter 3: Results -- Chapter 4: Discussion -- References

Notes

A thesis submitted in partial fulfillment of the degree of Master of Research.

Awarding Institution

Macquarie University

Degree Type

Thesis MRes

Degree

Thesis (MRes), Macquarie University, Faculty of Science and Engineering, 2021

Department, Centre or School

Department of Molecular Sciences

Year of Award

2021

Principal Supervisor

Paul R. Jaschke

Rights

Copyright: Pamela Rea Tsoumbris Copyright disclaimer: https://www.mq.edu.au/copyright-disclaimer

Language

English

Extent

61 pages

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