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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 <> by modifying the contents of existing exosomes through the use of the protein tag 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.

Over the past few months we have:

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.