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 <> 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.  
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Pharmaceutical companies rely on various non-human model systems to test the efficacy and toxicity of drug candidates in development. However, these systems may not be predictive of drug behavior in humans. To better predict drug behavior in human trials, a synthetic model that more closely mimics ''in vivo'' drug response is desirable.  Better ''in vitro'' predictions of drug toxicity and efficacy may lead to safer, more effective therapies.
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Over the past few months we have:
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One promising model under development is the introduction of genetic circuits to populations of cells to produce organoids. These synthetic systems are compositionally similar to organs and respond to external stimuli in a comparable manner. The formation and maintenance of these structures requires coordinated behavior between individual cells based on their local context. As a means to coordinating behavior, the 2013 MIT iGEM team is developing an exosome mediated cell-cell communication system for use in mammalian cells.
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Demonstrated Acyl-TyA targeting proteins to the cell membrane and into exosomes
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Designed a number of reporter constructs to assay for our signals:
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rtTA3
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Our exosome communication system employs two complementary signaling strategies. We are engineering sender and receiver cell circuits for testing signals including miRNA, recombinases, DNA-binding proteins, RNA-binding proteins, and proteases. We are particularly excited about the possibility of multiplexed communication using an exosomally delivered Cas9-CRISPR system.
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L7Ae
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Cas9-VP16
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Cas9-Split Venus
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We believe this method can be employed as a generalizable platform for intercellular communication. In concert with other synthetic biology modules, this work may be used in the future for creating mammalian systems that perform distributed computing, undergo multistep differentiation, or form complex microstructures.
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Cre Recombinase
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Designed Acyl-TyA fusion proteins with our signals:
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rtTA3
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L7Ae
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Cas9-VP16
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Cas9-Split Venus
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Cre Recombinase
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Demonstrated native exosomal microRNA repression with isolated exosomes and Jukat/HEK293 coculture experiments.
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Demonstrated activation of a reporter using the trans activator Cas9-VP16.
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Demonstrated DNA sensing using Cas9-Split Venus reconstitution.  
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Revision as of 02:32, 28 September 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 <> 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.

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