INFORMAZIONI SU
FRONZA Filippo
Supervisore: Prof. Schneider Green Diagnostics Lab — CRISPR based diagnostics |
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The project aims at the development of a CRISPR-based diagnostic test for pathogens, to be used in the context of a one-pot diagnosis of SARS-CoV2 and other influenza-line pathogens. Our approach involves the use of Cas9 endonuclease and of reprogrammed tracrRNA — called Rptr. The Rptr captures a matching RNA fragment and turns it into a non-canonical crRNA. The complex formed can then be used to selectively cleave a DNA reporter. This process would allow a fast and extremely precise detection of RNA sequences, which discriminate differences of a single base, in a shorter time than by PCR and without the need for costly or complex machinery and skilled healthcare practitioner. As part of the project, we aim at increasing the speed and sensitivity of the test by implementing Cas9 turnover, introducing the use of fluorescent probes in Rptr based technique, and developing a multiplexing test that will discriminate different pathogens and variants. |