We successfully designed and cloned a biobrick coding for the secretion of rat proNGF in pET43.1a and iGEM plasmid backbone pSB1C3, creating a new part Bba_K2616000 . We confirmed the the genetic construct by sequencing.

We confirmed the production of proNGF in three different ways:

• SDS PAGE followed by western blot.
• Mass spectrometry.
• Activity on the growth of rat E18 cortical cells.

We designed self-made microfluidic device in order to implement our final proof of concept.

We grew neurons on our self-made microfluidic chips and successfully observed axon growth in the presence of commercial NGF.

We successfully cloned a part coding for toxin/antitoxin (CcdB/CcdA) system in iGEM plasmid backbone, creating a new part Bba_K2616002

We observed survival and normal growth of our engineered bacteria at 25°C and 37°C and absence of growth at 18°C and 20°C, showing the efficiency of the kill switch if our bacteria are released in the environment.

In search of a biocompatible conductive polymer to confine bacteria, we successfully polymerized PEDOT:Ts and PEDOT:Cl on alumina oxyde membrane filters. We also partially polymerized PEDOT:PSS.

We demonstrated that a polymer-coating enhances the properties of the membranes as PEDOT:Ts-coated and PEDOT:Cl-coated membranes are more conductive than other membranes. Moreover, experiments showed a better biocompatibility for the polymer-coated membranes compared to the gold-coated ones.

We succeeded in finding appropriate membranes for our project, namely PEDOT:Ts and PEDOT:Cl. We would like to do further research and improve the way we polymerized PEDOT:PSS because it is widely used in organic electronic.