My First AIRC GRANT 2023

Cancer cells often show reduced DNA repair capacity and higher genetic instability. Also the efficacy of chemotherapy treatments is strongly influenced by cellular DNA repair capacity. Although our understanding of DNA repair mechanisms has grown rapidly, toolkit for studying DNA repair enzymes has lagged behind. Standard cell-based assays for monitoring repair activities are well-established technologies but they are also indirect, laborious, and with limited applicability for clinical applications.

The development of activity-based sensing platforms for the monitoring of repair enzymes, and their translation into clinical settings, can pave the way for improved early cancer diagnosis and novel personalized therapies based on repair capacity assessments in patients. To achieve this objective, CRISPR-based technology appears particularly attractive because it can be easily harnessed to convert the readout to a pointof-care tool. CRISPR also demonstrated the capacity to work robustly using biological samples and clinical specimens, and showed potential for high throughput screening (HTS) applications.

The transformative goal of my research program is to develop a synthetic biology toolkit enabling real-time, activitybased monitoring of DNA glycosylases involved in base excision repair (BER) of base pairs containing 8-oxoG. My efforts will be mainly devoted to the development of an activity-based CRISPR platform for the realtime monitoring of MutY homolog DNA glycosylase (MUTYH). The platform will be further adapted to produce innovative HTS assays (CRISPR-HTS) of small molecule modulators of MUTYH activity.