Modular Construction of Bacteriophage for Diagnostic Systems
Introduction and Aim
Personal medicine and non-invasive biological measurements are the fastest growing areas in biotechnology. Viruses and bacteriophages are central to a number of biotechnology applications spanning diverse fields, from adenovirus for gene therapy in humans to M13 phages for use in lasers and batteries. These simple organisms have also provided us with much of our understanding of gene regulation and replication. The specific interaction that enables a virus to infect a host and carry out its multiplication process is at the heart of many new emerging technologies that have the capacity of providing breakthroughs in multiple fields of research.
Lambda phage is a well-characterised bacteriophage where all steps in the infection and life cycles are known. Multiple biotechnological tools based on it have also been developed (such as recombineering and Gateway® cloning). Our aim is to use the lambda bacteriophage to explore the avenue of producing a modular chassis. For example, broad host phage could be used for biosensing and biotechnological applications and high specificity could be explored for therapy. Host broadening and host reassignment can both (and have been shown to) be achievable by fusing genes from related phages (T4 / lambda) and by directed evolution.
Experimental Approach
Using available data on how different phages infect cells and provided the time restrictions, we will assess the modularity of the infection strategies as follows:
• Bioinformatics analysis of key lambda genes involved in host specificity and infection mechanism will be used to analyse relating phylogeny to known mechanisms, to explore modularity / host reassignment or broadening host range.
• Host specificity factors (i.e. tail fibre protein [stf gene, loci:lambdap27/lambdap28] and tip attachment protein J [J gene, locus: lambdap21]) will be targeted for engineering with a view to increase host infectivity (against a LamB / OmpC E. coli negative mutants) or to alter host specificity (against a second type species – potentially, Photorhabdus luminescens or a Rhizobium type strain as target).
• Engineering of phage cycle regulators will be performed to allow exogenous control of lytic / lysogenic cycle and to allow introduction of a biosensor.
Impact of Findings
We hope to establish lambda phage modularity as a proof-of-principle showing that phages can be engineered as biosensors that are cheap to make, stable for transportation and not requiring specialist knowledge to use. We see this as a potential route towards developing affordable point-of-care assays for pathogen detection.
Previous efforts so far have yielded great results regarding the proof of concept for quickly detecting bacteria using phages, and we would like to take the technology one step further by producing a single robust lambda phage-based multi-purpose chassis.
What should this page contain?
- A clear and concise description of your project.
- A detailed explanation of why your team chose to work on this particular project.
- References and sources to document your research.
- Use illustrations and other visual resources to explain your project.
Advice on writing your Project Description
We encourage you to put up a lot of information and content on your wiki, but we also encourage you to include summaries as much as possible. If you think of the sections in your project description as the sections in a publication, you should try to be consist, accurate and unambiguous in your achievements.
Judges like to read your wiki and know exactly what you have achieved. This is how you should think about these sections; from the point of view of the judge evaluating you at the end of the year.
References
iGEM teams are encouraged to record references you use during the course of your research. They should be posted somewhere on your wiki so that judges and other visitors can see how you though about your project and what works inspired you.
Inspiration
See how other teams have described and presented their projects:
