Til bevillingsoversigt

Sensing Genomic Sequences using the Bacterial CRISPR Immune System: a Potential New Sensor System

Carlsbergfondets internationaliseringsstipendier

What

I will examine how the bacterial immune system (CRISPR/Cas9) can be adapted to a versatile sensor system that generates a color visible to the naked eye. Such a sensor system could easily be adapted to detecting any given DNA/RNA sequence and it will be able to combine the specificity and sensitivity from current state of the art machinery. Such a sensor system would make it possible to test for infections or genomic mutations at remote locations or in our homes. Such a sensor system would have the potential to revolutionize diagnosis, sanitary control, food security and other areas where polynucleotide sequence detection is essential.

Why

To accommodate the increasing global population with an expected increased international mobility, we must go beyond what is currently possible and develop new tools for rapid and sensitive diagnosis, sanitary control and food security. Such novel tools are required to be robust and easy to use, while still possessing the sensitivity known from current technologies. We aim to develop a sensor technology, which will be able to detect targeted polynucleotide sequences within a few minutes by adding a swap with the sample to a tube. If the target sequence is in the sample, the liquid in tube will generate a clear color that can be detected by the naked eye.

How

The sensor system will have two components (hybrid-proteins) each consisting of two components: a nuclease deficient Cas9 (dCas9) and an enzyme, Horseradish Peroxidase, split into two parts. dCas9 can be programmed into specifically binding the target DNA and the Horseradish Peroxidase will facilitate a sensitive readout. If the target sequence is present in a sample, Horseradish Peroxidase will generate a color upon recognition. The sensor system will be developed and examined using target DNA sequences for specific detection of the virus causing HPV, but the sensor system can be adjusted to detect a variety of bacterial and eukaryotic microbes.

SSR

The proposed sensor technology will have a cascade of applications in fundamental and applied science. Such applications include point-of-care diagnostics in resource limiting areas, sanitary control in hospitals and in food production. The development of the proposed technology could be central to accommodate an expected increased pressure on our healthcare, food production and sanitary control.