Building a library of specialised killer phages
Lead Research Organisation:
University of Warwick
Department Name: Warwick Medical School
Abstract
Programme overview:
This MRC-funded doctoral training partnership (DTP) brings together cutting-edge molecular and analytical sciences with innovative computational approaches in data analysis to enable students to address hypothesis-led biomedical research questions. This is a 4-year programme whose first year involves a series of taught modules and two laboratory-based research projects that lead to an MSc in Interdisciplinary Biomedical Research. The first two terms consist of a selection of taught modules that allow students to gain a solid grounding in multidisciplinary science. Students also attend a series of masterclasses led by academic and industry experts in areas of molecular, cellular and tissue dynamics, microbiology and infection, applied biomedical technologies and artificial intelligence and data science. During the third and summer terms students conduct two eleven-week research projects in labs of their choice.
Project:
Infectious diseases are one of the leading cause of human morbidity and mortality. With the onset of the antibiotic era bacterial infections became significantly less threatening. However, bacteria have developed resistance to antibiotics over time. Since the rate of discovery of new antibiotics is slow, bacterial infections are becoming more threatening. Although the current viral epidemic has rightly taken the spotlight of government, scientists, industry and public health officials, the long term threat of antimicrobial resistance remains.
One way of treating bacterial infections is by phage therapy rather than a drug. This method has proven more effective and with a higher therapeutic index. Phage therapy is adaptable as phages evolve naturally to fit their target. However a key problem is that there are not enough phages able to specifically target all the various harmful bacteria.
For this reason, this PhD project work will focus on engineering a phage towards new therapeutic purposes (i.e. vaccines) and towards expanding their antimicrobial activity to non-permissive hosts. The project will focus on improving the existent methods to engineer phages, followed by optimising the upstream and downstream bioprocessing of respective engineered phage for in vivo trials. The end goal is to improve the phage therapeutic landscape and improve phage engineering techniques for future research.
This MRC-funded doctoral training partnership (DTP) brings together cutting-edge molecular and analytical sciences with innovative computational approaches in data analysis to enable students to address hypothesis-led biomedical research questions. This is a 4-year programme whose first year involves a series of taught modules and two laboratory-based research projects that lead to an MSc in Interdisciplinary Biomedical Research. The first two terms consist of a selection of taught modules that allow students to gain a solid grounding in multidisciplinary science. Students also attend a series of masterclasses led by academic and industry experts in areas of molecular, cellular and tissue dynamics, microbiology and infection, applied biomedical technologies and artificial intelligence and data science. During the third and summer terms students conduct two eleven-week research projects in labs of their choice.
Project:
Infectious diseases are one of the leading cause of human morbidity and mortality. With the onset of the antibiotic era bacterial infections became significantly less threatening. However, bacteria have developed resistance to antibiotics over time. Since the rate of discovery of new antibiotics is slow, bacterial infections are becoming more threatening. Although the current viral epidemic has rightly taken the spotlight of government, scientists, industry and public health officials, the long term threat of antimicrobial resistance remains.
One way of treating bacterial infections is by phage therapy rather than a drug. This method has proven more effective and with a higher therapeutic index. Phage therapy is adaptable as phages evolve naturally to fit their target. However a key problem is that there are not enough phages able to specifically target all the various harmful bacteria.
For this reason, this PhD project work will focus on engineering a phage towards new therapeutic purposes (i.e. vaccines) and towards expanding their antimicrobial activity to non-permissive hosts. The project will focus on improving the existent methods to engineer phages, followed by optimising the upstream and downstream bioprocessing of respective engineered phage for in vivo trials. The end goal is to improve the phage therapeutic landscape and improve phage engineering techniques for future research.
| Title | Lambda red recombineering in P2 bacteriophage |
| Description | In vivo recombineering is commonly catalysed by bacteriophage-encoded homologous recombination systems, such as the coliphage ? Red system (Reference: Shyam Sharan et. al., PMC 2010). Since induction of the phage ? in the presence of P2 prophage causes inhibition of protein synthesis leading to cell death (Reference: Gunnar Lindahl et. al., PNAS 1970), recombineering in P2 lysogens was overlooked until recently, with most of the work being performed in the lytic state. Although there are standard methods for engineering bacteriophages in a lytic state, they take a longer time at best to generate and screen for recombinants. Tackling the P2- ? interference mechanism while at the same time maintaining the ? recombineering attributes was therefore crucial for engineering mutants of bacteriophage P2. We found that a combination of recA, lexA host, del1old prophage and the pORTMAGE plasmid (Nyerges et al., 2016), can successfully delete genetic material and insert instead a selection marker to facilitate screening for the successful recombinant mutants. We also demonstrated scarless editing of the prophage via the same approach, by following the MAGE workflow (Reference: R. Gallagher et. al., 2014 Nature America Inc) and using ssDNA as recombineering substrate. |
| Type Of Material | Technology assay or reagent |
| Year Produced | 2020 |
| Provided To Others? | No |
| Impact | The approach is used directly in my current research. |
| Description | Evaluation of engineered bacteriophage phage-system as an antimicrobial and vaccine agent |
| Organisation | GlaxoSmithKline (GSK) |
| Country | Global |
| Sector | Private |
| PI Contribution | We designed a method to produce pure high titer transducing particles containing confidential GSK cargo. We also built the cargo and demonstrated its activity in vitro. My direct contribution here was the downstream processing of the particles to be further used in mice trials. |
| Collaborator Contribution | GSK designed and provided the cargo parts. They are also to perform in vivo mice experiments in May 2020. |
| Impact | Scientific Paper due. |
| Start Year | 2018 |
| Description | Family day outreach Phage hunters stall |
| Form Of Engagement Activity | Participation in an open day or visit at my research institution |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Public/other audiences |
| Results and Impact | This was a family day event for inspiring children to pursue a career in science. My part was to describe in general terms to the parents and kids what a bacteriophage is and to teach children how to build a model of a bacteriophage from paper, clay and sticks. |
| Year(s) Of Engagement Activity | 2019 |