Team:MIT/Protein

From 2013.igem.org

iGEM 2012

Overview

  • Project Overview

miRNA Signal

  • Overview
  • siRNA Characterization
  • Exosome Isolation and Co-Culturing
  • Cell-Cell Co-Culturing

Protein Signals

  • Overview
  • GFP
  • rtTA3
  • Cre
  • L7Ae
  • Cas9-VP16

Novel DNA Sensor: Cas9 Split Venus Fusion

  • Overview
  • Leucine Zipper Fusion
  • DNA Sensing

Our BioBricks

  • Favorites
  • All BioBricks

Attributions

  • Attributions

Overview of Protein-based Cell-Cell Communication

While miRNA are useful in their own respect, it’s not currently known how miR-451 is naturally targeted into exosomes, making it difficult to engineer a system of sending any generic RNA. Luckily, research has shown that proteins can be targeted into exosomes by using an oligomerizing protein domain called Acyl-TyA (Shen, 2011). By fusing our protein signal to an Acyl-TyA domain, it’s possible to target our signal protein into exosomes and send it to another cell to evoke a response.

Our goal is to use Acyl-TyA to target generic protein signals into exosomes which allow for cell-cell communication by facilitating the transfer of a signal protein from a sender cell to a receiver cell. The general procedure begins with creating an Acyl-TyA fusion protein and testing the signal’s function within single cells to determine whether the fusion was successful. Then, once the signal’s function is verified, we use Jurkat T cells to produce exosomes containing our signal and apply these exosomes to receiver cells—the signal would then enter the receiver cells via the exosomes and activate a specific receiver circuit. Finally, the signal producing Jurkat T cells are to be cocultured together with receiver cells to determine whether cell-cell communication has been achieved.

Shen, B et al. Protein targeting to exosomes/microvesicles by plasma membrane anchors. J Biol Chem. (2011)