Exosome Mediated Mammalian Cell-Cell Communication

Overview

miRNA Signal

Protein Signals

Novel DNA Sensor: Cas9 Split Venus Fusion

Our BioBricks

Attributions

The MIT iGEM team sought to create a new mode of engineered intercellular communication for use in synthetic biology by modifying the contents of existing exosomes through the use of naturally occurring miRNA and the protein domain Acyl-TyA. We built on existing research targeting proteins to exosomes to enable intercellular communication by targeting signal proteins into exosomes and into HEK 293 receiver cells.

Exosomal Cell-Cell Communication with miRNA

Jurkat T cells are known to produce a large number of exosomes which naturally contain high levels of miRNA 451. Using this natural system, our initial goal is to create a miRNA 451/Exosome sensor to begin our work with Exosomal communication.

Demonstrated Acyl-TyA targeting proteins to the cell membrane and into exosomes

Designed a number of reporter constructs to assay for our signals:

• rtTA3
• L7Ae
• Cas9-VP16
• Cas9-Split Venus
• Cre Recombinase

Designed Acyl-TyA fusion proteins with our signals:

• rtTA3
• L7Ae
• Cas9-VP16
• Cas9-Split Venus
• Cre Recombinase

Demonstrated native exosomal microRNA repression with isolated exosomes and Jukat/HEK293 coculture experiments.

Demonstrated activation of a reporter using the trans activator Cas9-VP16.

Demonstrated DNA sensing using Cas9-Split Venus reconstitution.