Part:BBa_K1179025

CAG_L7Ae

Constitutive expression of L7aE which is used as a translational repressor.

Overview

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.

L7Ae 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

Acyl-TyA-L7Ae: Exosome Isolation and Co-Culturing

Acyl-TyA-L7Ae: Cell-Cell Co-Culturing

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 1730
  Illegal XbaI site found at 1634
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1730
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1730
  Illegal XhoI site found at 12
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1730
  Illegal XbaI site found at 1634
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1730
  Illegal XbaI site found at 1634
  Illegal NgoMIV site found at 1322
  Illegal NgoMIV site found at 1479
  
• 1000
  COMPATIBLE WITH RFC[1000]