Team:MIT/Project

From 2013.igem.org

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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 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. <br><br>
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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. <br><br>
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Over the past few months we have:<br><br>
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<h3>Exosomal Cell-Cell Communication with miRNA</h3>
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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.
<li>Demonstrated Acyl-TyA targeting proteins to the cell membrane and into exosomes
<li>Demonstrated Acyl-TyA targeting proteins to the cell membrane and into exosomes

Revision as of 00:14, 26 October 2013

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

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.

    • Retrieved from "http://2013.igem.org/Team:MIT/Project"