Til bevillingsoversigt

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

Carlsbergfondets internationaliseringsstipendier

What

The project will investigate how the intriguing CRISPR/Cas9 system can be adapted to a versatile platform sensor. Such a sensor could easily be adapted to detect any given DNA/RNA sequence and it will be able to combine the specificity known from PCR with the ease-of-use known from ELISA-based tests. Once developed, the platform sensor has the potential to revolutionise 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 novel, rapid and sensitive tools for 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 platform technology, which will be able to detect targeted polynucleotide sequences and the readout is expected to be adaptable into a stick-dip test similar to well-known pregnancy test.

How

The platform sensor will have two components (hybrid-proteins) each consisting of three elements: Cell Penetrating Peptide (CPP), a nuclease deficient Cas9 (dCas9) and Protein-fragment Complementation Assays (PCA). CPP to facilitate crossing over the cellular membrane, dCas9 will be directed to target DNA using gRNA and PCA to facilitate a sensitive readout. If the target sequence is present in a sample, the two hybrid-proteins will generate a detectable signal upon recognition. The platform will be developed and examined using target DNA sequences for specific detection of the viruses causing HIV, HPV and herpes, but we expect the platform can be adjusted to detect a variety of bacterial and eukaryotic microbes.

SSR

The proposed platform 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.