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Revision as of 01:41, 2 November 2017

PhagED: a molecular toolkit to re-sensitise ESKAPE pathogens

Antibiotic revolution: a molecular toolkit to re-sensitise
Methycillin resistant S.aureus (MRSA) Vancomycin resistant S.aureus (VRSA) Klebsiella pneumoniae Pseudomonas aeruginosa Enterococcus faecalis/faecium Acinetobacter baumannii

Modular molecular toolkit for re-sensitisation of antibiotic-resistant pathogens using CRISPR delivered by a two-phage system.

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ABOUT PhagED

The threat posed by antibiotic resistant bacteria is a pressing issue which must be addressed. It is difficult and expensive to develop new antibiotics, so our project is designed to make currently available antibiotics useful again. Our aim is to create a toolkit to re-sensitise pathogens to antibiotics using CRISPR and a two-phage system, based on work by Yosef et al. (2015, doi:10.1073/pnas.1500107112). An engineered lysogenic phage will transfer a CRISPR system to its host bacterium, designed to cleave resistance genes and also confer protection from an engineered lytic phage. When this lytic phage is added to the population, it kills any bacteria that have not been successfully re-sensitised. We chose to target genes found in the highly-resistant ESKAPE pathogens, and worked with 4 different phages - P1, lambda, T4 and T7. Our system was modelled in silico and tested empirically on a specially designed E. coli testing platform.

Figure: Yosef et al., 2015

TAKE A CLOSER LOOK

Project

Get to know PhagED better.

Parts

Blocks of DNA that we've used.

Safety

Safety always comes first.

Human Practices

How does PhagED fit into the context of our lives?

Meet The Team

Filippo Abbondanza

MSc Synthetic Biology & Biotechnology

Engineering the lysogenic lambda phage with a CRISPR system to target resistance gene fragments.

Erin Corbett

MSc Synthetic Biology & Biotechnology

Engineering E. coli to create our mock pathogen testing platform, and engineering the lytic T7 phage.

Yunqi He

MSc Biochemistry

Engineering the lysogenic P1 phage with a CRISPR system to target resistance gene fragments.

Ti He

MSc Biotechnology

Engineering the lysogenic lambda phage with a CRISPR system to target resistance gene fragments.

Lydia Mapstone

MSc Synthetic Biology & Biotechnology

Engineering E. coli to create our mock pathogen testing platform and engineering the lytic T4 phage using BRED.

Yuri Matsueda

MSc Biotechnology

Engineering the lysogenic P1 phage with a CRISPR system to target resistance gene fragments.

Anton Puzorjov

MSc Bioinformatics

Building a model of bacteria-phage interactions in two-step re-sensitisation combining both lysogenic and lytic phages.

Yating Wang

MSc Drug Discovery & Translational Biology

Engineering the lysogenic phage P1 with a CRISPR system to target resistance gene fragments.


Owen Yeung

MSc Synthetic Biology & Biotechnology

Engineering the lysogenic lambda phage with a CRISPR system to target resistance gene fragments.

The Edinburgh_OG team members developed the iGEM project as part of our Masters dissertations at the University of Edinburgh. Due to academic requirements, the iGEM project, PhagED, was divided into individual sub-projects to allow the production of unique dissertations. Each of us completed a dissertation based on the work we did for iGEM, and these dissertations are available upon request.

Supervisors

Dr Elise Cachat

School of Biological Sciences

Academic supervisor.

Dr Heather Barker

School of Biological Sciences

Supervising lab work.

Holly Robertson-Dick

Industrial Liaison

Supervising iGEM administrative work.

Dr John White

School of Chemistry

Supervising phage work.

Dr Filippo Menolascina

School of Bioengineering

Supervising bacteria-phage modelling.

Dr Russell Brown

School of Biological Sciences

Supervising P1 phage construction.

Sponsors

We are very thankful to those who are helping us to make it happen.

FEEL FREE TO GET IN TOUCH

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