Synthetic biology approaches to probe neuronal and synaptic function

Research team

  • Simona Capsoni
    Professoressa su convenzione
  • Marco Mainardi
    Researcher
  • Paola Tognini
    Fellow
  • Jacob Ajesh
    PhD Student
  • Renata Cerna
    PhD Student
  • Valerio Corvaglia
    PhD Student
  • Lucia Rota
    PhD Student

In 2019, research concerning synthetic biology focused on the selection of nanobodies from the PISA platform and on the development of the PISA2.0 platform, in which the selection of post-translational modifications takes place through the expanded genetic code, i.e. by integrating tRNA-AAt/RNA synthetase pairs in the genome of yeast, for the incorporation of non natural amino acids. In particular, nanobodies against phosphorylated Tau were selected and subsequently validated in vitro and in cell cultures, and using the IACT nanobodies technique against Neurexin 3, Neuroligo 3, TDP43 and against CD19. A new selective and reversible degradation system of intracellular proteins has also been developed by combining nanobodies with photo-inducible (light sensitive) modules.
The activity related to the study of Tau functions also led to the development of a method to analyze the consequences of nuclear localization of the tau protein on gene expression, in particular of the VGLUT1 gene, and to the identification of an ERK kinase inhibitor as a potential anti-aggregating agent of the tau protein.
An in vitro setup was developed using the CST sensor for the identification of potential therapeutic molecules against Tau aggregation.
A transgenic model in zebrafish was also generated and characterized that expresses the CST sensor in neurons in order to validate an in vivo setup for the selection of anti-aggregation drugs.