A researcher uses an imager to examine results from a gel electrophoresis assay

Identifying Pathogenic Genetic Variants with Gene Editing

To study the disease-causing effects of genetic mutations in human cells, the precision gene editing tool CRISPR was used to introduce mutations into cells. High-throughput technologies were then used to examine the effects of these mutations on cells. Compelling evidence was produced that these techniques are effective in identifying well-known pathogenic mutations as well as predicting the likely pathogenicity of previously unknown mutations. 

DNA Mutations and Disease Susceptibility

DNA mutations can cause disease or affect an individual’s susceptibility to disease. DNA sequencing technologies have led to great advances in mapping the genetic code contained in DNA and also in identifying some of the genetic mutations that can cause problems when they occur in human cells. Despite huge advances in DNA sequencing technologies made in recent years, the significance of approximately 40% of genetic mutations listed within public databases remains unknown. In the context of COVID-19, there is a deficit of knowledge regarding the potential effect of mutations in the human receptor for SARS-CoV-2, the virus that causes COVID-19, on how susceptible a person is to viral infection and their clinical response once infected.

Testing Techniques to Identify Pathogenic Mutations

The consortium of researchers involved in this project used a precision gene editing tool called CRISPR to introduce thousands of genetic mutations into human cells. They used high-throughput technologies to simultaneously examine thousands of gene mutations and their potential to cause disease. Using this technique allows for vast amounts of scientific data to be collected in a single experiment. The ultimate goal of this consortium is to utilise these approaches to examine how genetic elements influence SARS-CoV-2 infection susceptibility. In the first instance however, they tested the techniques using an established cancer cell model, which allowed for faster troubleshooting.

The research team successfully utilised the technologies described to predict the disease-causing potential of genetic mutations in the cancer-associated gene EGFR. Additionally, the treatment of these cells with well-known drugs designed to treat cancer, enabled the identification of new mutations responsible for drug resistance.

Value and Future Potential of the Project’s Results

The results produced in this project provide compelling evidence that the techniques developed and tested can be used to identify well-known pathogenic mutations as well as to predict the likely disease-causing potential of previously unknown mutations. The researchers are in the process of conducting further research to validate their findings.

In the future, the efforts of this research consortium have the potential to help identify patients who are less likely to respond to certain treatments based on their genetic profiles and to inform the establishment of patient-specific courses of treatment.

Banner image above: A researcher uses an imager to examine results from a gel electrophoresis assay.

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Lead Researchers