"This proposal is intended to develop biosensors, named CDKACT and CDKCONF, as smart tools to detect and study key cancer biomarkers, such as CDK4 and CDK5 kinases, which may be of potential value for cancer diagnostic and theragnostic purposes, as well as to propose new selective compounds for cancer therapeutics by screening for non-ATP-competitive inhibitors of these targets.This is a multidisciplinary project at the interface between chemistry, biochemistry, biology, and cell and animal imaging technologies. A wide variety of state-of-the-art technologies will be applied, including: (i) innovative chemical approaches for the design, synthesis and engineering of peptide and protein biosensors, (ii) biochemical, biophysical, and fluorescence-based approaches for characterization of the sensitivity, specificity and physicochemical features of the biosensors, (iii) cell biology and fluorescence imaging approaches for characterization and validation of these biosensors in cellulo and in vivo, (iv) application of fluorescent biosensors for development of diagnostic assays, and (v) application of fluorescent biosensors for development of high throughput screening assays and identification of selective non-ATP competitive inhibitors for cancer therapeutics.This program will have a large scientific, technological and economic impact, by providing new diagnostic tools and molecules with selective inhibition profiles with application to life sciences and biomedicine. Expected results will contribute to improve early detection of alterations associated with cancer cell proliferation, and will provide novel kinase inhibitors for cancer therapy."