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Revision as of 14:28, 16 October 2018
Overview
Welcome to the Nottingham iGEM 2018 Human Practices pages. This overview will introduce you to the projects that we investigation for our Human Practices efforts, as well as why we investigated them and what our results showed.
For our silver medal criteria, we complied 3 reports on the impact of our project, how people feel about phage therapy and what current legislation there is on phage therapy. The objective of compiling these reports was to investigate whether our work was responsible and good for the world. We engaged with a variety of communities including field experts, community discussion groups and scientific articles and research papers to gain a wide scope of information to demonstrate the positive impact of our work.
For our gold medal criteria, we analysed how we could integrate the findings of our discussion group and field expert interviews into our project. We chose to document this via a clear flowchart so that we can explain how we have taken on board feedback and modified our project as a result. We hope enjoy learning about our Human Practices work on this year’s project. Click the dropdown menu and pages to find our more details about our work!
Silver
Gold
For our gold medal criteria, we analysed how we could integrate the findings of our discussion group and field expert interviews into our project. We chose to document this via a clear flowchart so that we can explain how we have taken on board feedback and modified our project as a result. We hope enjoy learning about our Human Practices work on this year’s project. Click the dropdown menu and pages to find our more details about our work!
Safety and Ethics
Lab photos
Project description
We are planning to use dCas9 and asRNA to silence the production of toxins A and B in C. difficile. C.difficile is known for causing hospital-acquired diarrhoea in the Western world. We want to prove a concept of asRNA and/or dCas9 as an alternative to antibiotic treatment against C.difficile. As a result, we will neutralise the negative impacts of bacteria on human health rather than completely killing it. We will create and test a promoter library in both E. coli and C. difficile, express anti-sense RNAs in C. difficile, express dcas9/sgRNA in E. coli and C. difficile. Dcas9 is planned to be used to repress the reporter gene gusA, by dcas9 binding to the toxin promoters placed upstream of gusA, in E. coli and C. difficile. If this repression is successful the system will be used to repress the C. difficile toxin genes.
Whole organisms
Species used as a chasis
Parts
See our parts page
Experiments undertaken with organisms and parts
All parts will be used to create and test a promoter library in both E. coli and C. difficile, express asRNAs in C. difficile, express dcas9/sgRNA in E. coli and C. difficile. The asRNAs will be used to repress the expression of toxin A and B in C. difficile. Dcas9 will be used to repress the reporter gene gusA (cas9 binding to the toxin promoters placed upstream of gusA) in E. coli and C. difficile. If this repression is successful the system will be used to repress the C. difficile toxin genes.
If toxin supression in E.coli is successful, we plan to use E.coli sexpress (NEB express + R702) to conjugate E.coli containing asRNA or dCas9 vector into C.difficile 1801 to test toxin supression in the host organism. Given enough time, we plan to insert asRNA/dCas9 system into bacteriophage specific to C.difficile 1801 (ribotype 078) as an ultimate test of our project efficacy.
How real people would use our product
Out product would be used in the human body, or in food (Examples: anti-cancer bacteria, bread made with engineered yeast, engineered rice plants)
Risks associated with real use
Our project is basically focused on proving the concept that one can use bacteriophage therapy against C.difficile. We wanted to explore alternative treatment options due to the increasing problem of antibiotic resistance. Of course, we do not claim in any way that our final product will be straight away ready for therapeutic use. In the best scenario, it would need to be tested in vivo in preclinical studies and in clinical trials to assess their safety and test for any side effects. This would be done with ethical approval. Pharmacokinetic studies and biochemical experiments would be carried out to determine optimum conditions for use.
Our project is focused on using phage. Even though it is specific to C.difficile, its specificity may vary depending on different conditions. More research would be needed to investigate possible non-specific infection.
How experts have overseen our project to manage identified risk
Michelle Kelly, one of the safety officers at the lab our team has been working in, carried out a full laboratory safety induction and introduced the risk assessments and standard operating procedures that are adhered to when working in a Category 2 laboratory. She has also set up training records for each team member which requires signing off by advisors when each lab skill has been efficiently performed. She has been working on C.difficile for more than 5 years and possesses deep knowledge into proper project design that ensures safety of the researcher.
Nigel Minton (PI) has worked on C. difficile and Clostridia for over 20 years and has overseen the project development. Four other advisors who will also oversee the work that is completed, all of whom have worked on Clostridia for at least 3 years. These researchers will be responsible for ensuring SOPs and RAs are followed on a day to day basis. All parts of our project fall under university biosafety regulations as was checked by PI and safety officer.
Rules and guidance cover that helped manage identified risks
Safety training
Our team recieved security and safety training
Topics learnt about
Areas where we handled biological materials
Biosafety level of lab
How rules, training, containment and other procedures helped manage risks identified:
Each team member has completed a building safety induction and wet lab team members have completed a laboratory work induction. Each requires the successful passing of a quiz. Each lab team member has read and signed risk assessments for all of the work they will be completing and has a training record which has to be completed by a supervisor when training has been completed.
UK Regulations and their associated guidance are implemented, the University of Nottinghamhas defined appropriate standards for the growth, storage, disposal and transport of biological agents and genetically modified organisms within its premises. All these requirements are written down in the safety regulation forms. Our project was reviewed by the university safety committee to ensure that it follows the biosafety regulations of the laboratory we use.
All wet lab members were trained to work in group risk 2 laboratory and using the facilities. All regulations regarding waste disposal, equipment use and handling of dangerous chemicals were introduced by the safety officer. At least one supervisor was always present in the lab to ensure that the safety protocols are followed thoroughly.
In addition to standard procedures for working within a category 2 laboratory due to the nature of C. difficile no member of the team can enter the lab when they are taking antibiotics.
Compliance
Releasing organisms from our project
We will not be realsing any organisms from our project.
Gene drives
Our project does not use gene drives.
Resistance factors
We have used resistance factors, and have thus submitted an iGem check-in form.
Use of animals
We do not use any animals in our project.
Parts not on the whitelist
We have used parts not on the whitelist, and have therefore submitted an iGem check-in form.
Parts obtained outside of lab
We don't plan on using any parts outside of the lab.
