Alessandro Noto Email and Phone Number

My work is focusing on the role of WRN as a target of synthetic lethality in cells with the MSI phenotype. The study is based on the use of laboratory-generated biotinylation constructs that allow the analysis of short-term interaction kinetics.

Laboratory activity was focused on the study of the interaction between the helicase protein WRN and the ssDNAbp RPA in response to replicative stress induced by hydroxyurea. Through the use of immortalized cancer cells and fibroblasts from Werner Syndrome patients, I explored the importance of this interaction in maintaining genomic integrity. Study was performed via different techniques:-Bacterial Cultures-Cell cloning (Generation of new cell line models)-Viral… Show more Laboratory activity was focused on the study of the interaction between the helicase protein WRN and the ssDNAbp RPA in response to replicative stress induced by hydroxyurea. Through the use of immortalized cancer cells and fibroblasts from Werner Syndrome patients, I explored the importance of this interaction in maintaining genomic integrity. Study was performed via different techniques:-Bacterial Cultures-Cell cloning (Generation of new cell line models)-Viral infection-Generation of in vitro fragments-In vitro kinase assay-Pulldown assay-Use of siRNA-Immunofluorescence assay-Immunoprecipitation assay-Western Blot assay-Chromatin isolation-In situ Proximity Ligation Assay (PLA)-DNA fiber assay-Comet assay-MTT assay-Biotinylated proteins purification-Data analysis software usage (Microsoft Excel, ImageJ, Graphpad Prism, ImageLab, CometScore, Snapgene) Show less

My activity was focused on evaluating the anticancer effects of the combination of IFN-α and Romidepsin and establishing the mechanisms underlying their activity on melanoma cells and on the induction of the immune response (PD-1/PD-L1 signaling). Experiments were conducted on immortalized and patient metastatic melanoma lines, with and without mutation in the BRAF protooncogene. Study was performed via a number of techniques including:-Cell cultivation;-MTS assay;-Apoptosis… Show more My activity was focused on evaluating the anticancer effects of the combination of IFN-α and Romidepsin and establishing the mechanisms underlying their activity on melanoma cells and on the induction of the immune response (PD-1/PD-L1 signaling). Experiments were conducted on immortalized and patient metastatic melanoma lines, with and without mutation in the BRAF protooncogene. Study was performed via a number of techniques including:-Cell cultivation;-MTS assay;-Apoptosis assay;-RNA extraction-Real-time PCR-Western Blotting-Chromatin Immunoprecipitation-Phagocytosis Assays (Flow Cytometry)-Generation of IL-4-DC dendritic cells Show less