Refactoring bacteriophage as repurposed nanomachines
Lead Research Organisation:
Birkbeck, University of London
Department Name: Biological Sciences
Abstract
Strategic Research Priority: World Class Bioscience
Abstract
Bacteriophages are natural nanomachines, efficiently delivering DNA payloads into bacterial cells. Phage such as lambda are also general cloning vehicles, in which recombinant cargoes are packaged and delivered via natural virulence mechanisms. The targeting mechanisms of phages have been shown to be genetically malleable enough to target new cell surface proteins on their natural host[1]. Alternate strains of a given microbe can be accessed by the same phage machinery via engineering of the receptor binding protein[2,3]. The natural mechanism could in principle therefore be engineered to deliver a variety of synthetic payloads to a diverse range of target cell types.
Project
The aim of this project is to take the principles of variant payload and cell targeting, and refactor this via implementation of synthetic biology. We aim to demonstrate that the payload can be radically refactored to deliver new genes to targeted cells for a variety of end purposes, such as metabolic modification or more efficient killing of targets. Furthermore, we aim to discover whether cellular targeting can be made cross-species, as opposed to simply: receptor to receptor within a strain, or strain to strain within a species.
We propose to repurpose bacteriophage lambda, a system that has been extensively tailored for recombinant engineering, in order to deliver test payloads: for example Green Fluorescent Protein, or the DNA gyrase toxin CcdB. We additionally propose to engineer the E.coli LamB targeting protein[1,4] of lambda to infect other gram negative bacteria, e.g. nosocomial pathogens such as P.aeruginosa, and S.marcescens. We also suggest similarly repurposing a gram positive host infecting phage such as SPP1[5], to potentially target pathogens such as S.aureus.
Abstract
Bacteriophages are natural nanomachines, efficiently delivering DNA payloads into bacterial cells. Phage such as lambda are also general cloning vehicles, in which recombinant cargoes are packaged and delivered via natural virulence mechanisms. The targeting mechanisms of phages have been shown to be genetically malleable enough to target new cell surface proteins on their natural host[1]. Alternate strains of a given microbe can be accessed by the same phage machinery via engineering of the receptor binding protein[2,3]. The natural mechanism could in principle therefore be engineered to deliver a variety of synthetic payloads to a diverse range of target cell types.
Project
The aim of this project is to take the principles of variant payload and cell targeting, and refactor this via implementation of synthetic biology. We aim to demonstrate that the payload can be radically refactored to deliver new genes to targeted cells for a variety of end purposes, such as metabolic modification or more efficient killing of targets. Furthermore, we aim to discover whether cellular targeting can be made cross-species, as opposed to simply: receptor to receptor within a strain, or strain to strain within a species.
We propose to repurpose bacteriophage lambda, a system that has been extensively tailored for recombinant engineering, in order to deliver test payloads: for example Green Fluorescent Protein, or the DNA gyrase toxin CcdB. We additionally propose to engineer the E.coli LamB targeting protein[1,4] of lambda to infect other gram negative bacteria, e.g. nosocomial pathogens such as P.aeruginosa, and S.marcescens. We also suggest similarly repurposing a gram positive host infecting phage such as SPP1[5], to potentially target pathogens such as S.aureus.
Studentship Projects
| Project Reference | Relationship | Related To | Start | End | Student Name |
|---|---|---|---|---|---|
| BB/M009513/1 | 01/10/2015 | 31/03/2024 | |||
| 1627421 | Studentship | BB/M009513/1 | 01/10/2015 | 30/09/2019 | Hugo Villanueva |
| Description | We've been able to discover 3 different structures associated in viruses that attack bacteria. Using these structures, we want to create mutations to create mutant viruses. Normally these viruses attack non-pathogenic bacteria and we want to redirect them to pathogenic bacteria. |
| Exploitation Route | Hopefully, someone is able to make the hybrid phage in vivo. |
| Sectors | Agriculture, Food and Drink,Healthcare,Pharmaceuticals and Medical Biotechnology |