General Lab Safety
Before we started our project, the team was given a lab induction by Dr Matthew Peake, our Senior Biological Research Technician, which covered all biological and non-biological hazards within the lab. We were inducted on lab conduct and how to use laboratory facilities correctly. Whilst working in the labs, we were supervised at all time by academics and were not permitted to work out of hours. The use of specific equipment e.g ThermoFisher Plate Reader was inducted by the appropriate academic.
The lab has safety files that are revised and kept up to date and the appropriate forms are signed by the participating team member in the experiment. COSHH forms were filled out for every compound used in experiments and were updated when required.
As well as a lab induction, our facilities coordinator Leda Constantinides inducted us on emergency procedures and fire safety of the building. All our labs require smart card access, meaning only those qualified to be in the labs have access to them. This was arranged by Leda Constantinides, in which our cards only provide us with access to the labs from 9am-5pm.
In order to minimise the risks within labs, there are legislations in place within iGEM, the UK and worldwide. Our lab work is in line with these legislations. Each team member must be familiar with the policies set by Newcastle University and must abide to these policies. We have academic staff supporting our team to ensure the team follows and practices these policies. Those who do not abide to these policies be dealt with by an academic staff.
Aseptic technique was practiced to prevent biocontamination and unintended release of organisms within the laboratory and beyond. Blue lab coats were worn in the lab at all times and were not taken outside. White lab coats are provided for accessing different labs to prevent biocontamination. All waste is collected by the Central Services in the building which is incinerated or autoclaved. To minimise the risks of biocontamination and release of organisms, we chose non-pathogenic strains of organisms.
CRISPR-Cas13a SHERLOCK System
CRISPR-Cas9 for gene editing is the most commonly known CRISPR-Cas system, particularly due to the many ethical issues of this system prompting debates on the potential to enhance humans. Whilst CRISPR-Cas9 cuts and adds DNA, the CRISPR-Cas13a SHERLOCK system differs as Cas13a cuts RNA . SHERLOCK is a highly sensitive diagnostic system to detect diseases. Whilst researching the CRISPR-Cas13a system, the safety and ethical implications of the system has not been well studied so the safety of this system has not been determined. However, this system would not be used for gene editing and the safety measurements of other CRISPR-Cas systems involved in gene editing would not be applicable.
Biosensors using E. coli as a chassis
We attempted to introduce genes encoding fluorescent proteins into E. coli. We used E. coli DH5α for our biosensors, which is a non-pathogenic strain. In order to introduce genes into E. coli, we followed general molecular biology procedures such as cloning and transformation. Dependent on the plasmid we used, the selection of transformants involved the use of the antibiotics chloramphenicol, kanamycin and ampicillin. This is reported as a potential carcinogen so suitable protective equipment was worn when handling it.
Currently, our biosensors will not be implemented in the National Health Service (NHS) as such diagnostic tool would require vigorous testing. We do not have the appropriate permits from a governing body or proper supervision by a clinician. Ideally, we would like to test our product in a lab setting, however, if our product was used in the diagnosis of Parkinson’s Disease, it would likely undergo more vigorous testing than our labs allow.
If our project was to develop in the future, we would hope that the test be administered by healthcare professionals in the form of a blood test and skin swab (usually on the neck or face) and samples be analysed on a plate reader in healthcare laboratories.
As practical safety is an important feature of our tool, the ethical implications are just as important to us. At the start of the project, we had to think to ourselves, will introducing an early indicative tool be helpful to diagnose people with Parkinson's Disease?
Upon reading the NICE Guideline  on Parkinson’s Disease, it is clear that there are ethical issues we may encounter on the ethics of diagnosing Parkinson’s Disease in a non-clinical way.
5.10 “…Given these difficulties, it is generally accepted that the diagnosis of PD should be based on clinical findings. The most widely accepted clinical criteria for the diagnosis of PD are those introduced by the United Kingdom Parkinson’s Disease Society Brain Bank (Exhibit 5A).”
The guide also emphasised how important it is to make an accurate diagnosis in a person with suspected PD -
5.11 “It is important to make an accurate diagnosis in a person with suspected PD as this has an important bearing on prognosis”
Upon meeting with professionals in Parkinson's Disease and diagnostics, it was continuously emphasised that we need to ensure false positives were minimised to ensure the highest accuracy. Particularly with Parkinson's Disease, tools for indication or diagnostics must be highly accurate as this allows for the correct treatment options and misdiagnosis can cause emotional and physical stress.
- Cohen J. New CRISPR tool can detect tiny amounts of viruses. Science Magazine (Cited 20 October 2019). Available from: https://www.sciencemag.org/news/2017/04/new-crispr-tool-can-detect-tiny-amounts-viruses
- NICE. Parkinson's Disease: Full Guidline (Cited 20 October 2019). Available from: https://www.nice.org.uk/guidance/CG35/documents/parkinsons-disease-full-guideline-first-consultation2