Team:BIOTEC Dresden/Project v2

Temporal and spatial control of protein synthesis by in vitro recombination inside picoliter reactors

Manufacturing functionalized proteins in vitro poses a challenge, as it requires coordinated molecular assemblies and multi-step reactions. In this project we aim to control, over time and space, the production of proteins tagged with a silver-binding peptide for in situ silver nanoparticle nucleation inside microdroplets generated by microfluidic devices.

Combining a transcription-translation system with protein coding genes and a recombination logic inside microdroplets provides spatial control. Moreover, in the microfluidic chamber we can pinpoint the beginning of synthesis, and easily track and isolate the droplets. Site-specific recombination generates a molecular timer for temporal control of protein synthesis.

Unlike transcriptional regulation, this method gives true all-or-none induction due to covalent modification of DNA by Flp recombinase. Determining the transfer curve of inter-FRT site distance versus average recombination time allows the onset of gene expression to be predicted. We then apply this Flp reporter system as a powerful PoPS measurement device.

The project is split into three parts

Silver Nano-Particles

Here, it's attempted to create nanoparticles using a silver-binding peptide, as described in...

Gene Expression in Vesicles

Instead of gene expression in cells, it's attempted in vitro in vesicles.

Recombinase Pops Measurement Device

Utilizing a new biobrick, a recombinase mechanism is used to....