Team:MIT/L7Ae

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<h1>Overview</h1>
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L7Ae is an RNA binding protein that represses translation of the targeted transcript. Regulating gene expression on the translational level allows for faster repression of the desired construct than transcriptional repressors. L7Ae targets a specific sequence, called the 2x-kturn, on the 5’ end of the RNA. We use L7Ae to repress constitutive 2x-kturn_eGFP. L7Ae can be used to create more complex genetic circuits that are regulated on both the translational and transcriptional level. By targeting L7Ae to exosomes we create the opportunity for the creation of more intricate feedback looks in receiver cells.
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<b>Overview</b><br>
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<a href="#2">Characterization</a>
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<p>L7Ae is an RNA binding protein that represses translation of the targeted transcript. Regulating gene expression on the translational level allows for faster repression of the desired construct than transcriptional repressors. L7Ae targets a specific sequence, called the 2x-kturn, on the 5’ end of the RNA (Saito, 2010). We use L7Ae to repress constitutive 2x-kturn_eGFP. L7Ae can be used to create more complex genetic circuits that are regulated on both the translational and transcriptional level. By targeting L7Ae to exosomes we create the opportunity for the creation of more intricate feedback looks in receiver cells.</p>
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Saito, Hirohide. Synthetic translational regulation by an L7Ae–kink-turn RNP switch. Nat Chem Bio. (2010)
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<h1>L7Ae Characterization</h1>
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<h1>Characterization</h1>
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<a href="#1">Overview</a><br>
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<b>Characterization</b>
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<p>To demonstrate L7Ae mediated repression, we transfected constitutively L7Ae and constitutive 2x-kturn_eGFP. As we increased the amount of transfected L7Ae, we saw increased repression of GFP expression.*Note that there was a mistake made in this transfection. 5x the amount of GFP was transfected in the control well; however, L7Ae clearly represses eGFP expression</p>
<p>To demonstrate L7Ae mediated repression, we transfected constitutively L7Ae and constitutive 2x-kturn_eGFP. As we increased the amount of transfected L7Ae, we saw increased repression of GFP expression.*Note that there was a mistake made in this transfection. 5x the amount of GFP was transfected in the control well; however, L7Ae clearly represses eGFP expression</p>
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<div align="center"><img src="https://static.igem.org/mediawiki/2013/a/a5/L7aeone.png" width="200" height="250"></div>
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<img src="https://static.igem.org/mediawiki/2013/a/a5/L7aeone.png" width="200" height="250"></div>
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<h1>Acyl-TyA-L7Ae: Exosome Isolation and Co-Culturing</h1>
 
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<h1>Acyl-TyA-L7Ae: Cell-Cell Co-Culturing</h1>
 
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Latest revision as of 02:47, 29 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

Overview

Overview
Characterization

L7Ae is an RNA binding protein that represses translation of the targeted transcript. Regulating gene expression on the translational level allows for faster repression of the desired construct than transcriptional repressors. L7Ae targets a specific sequence, called the 2x-kturn, on the 5’ end of the RNA (Saito, 2010). We use L7Ae to repress constitutive 2x-kturn_eGFP. L7Ae can be used to create more complex genetic circuits that are regulated on both the translational and transcriptional level. By targeting L7Ae to exosomes we create the opportunity for the creation of more intricate feedback looks in receiver cells.

Saito, Hirohide. Synthetic translational regulation by an L7Ae–kink-turn RNP switch. Nat Chem Bio. (2010)

Characterization

Overview
Characterization

To demonstrate L7Ae mediated repression, we transfected constitutively L7Ae and constitutive 2x-kturn_eGFP. As we increased the amount of transfected L7Ae, we saw increased repression of GFP expression.*Note that there was a mistake made in this transfection. 5x the amount of GFP was transfected in the control well; however, L7Ae clearly represses eGFP expression

Retrieved from "http://2013.igem.org/Team:MIT/L7Ae"