Part:BBa_K1150025

uniCAS Red Light Switch Part I - Stimulator

uniCAS UV Light Switch Part II - Activator
Function	Cas9-protein linked to PIF6 induced gene expression control
Use in	Mammalians
RFC standard	RFC 25
Backbone	pSB1C3
Submitted by	[http://2013.igem.org/Team:Freiburg Freiburg 2013]

Introduction

Our construct contains the standardized version of Cas9 linked to PIF6. This device is one of two parts that you need to enable red-light induced gene targeting. Cotransfection of a PhyB+effector device, and target RNA allows you to recruit any desired effector to your desired Gene locus. Illumination with red light (660nm) will start the interaction and illumination with far red light (740nm) will stop the interaction. The Cas9-version used here is human codon optimized and the mutated version, unable to cleave DNA. PIF6 is human codon-ptimized.

References:

Muller, K.; Engesser, R.; Metzger, S.; Schulz, S.; Kampf, M. M.; Busacker, M. et al. ((Nucleid Acid Research; 2013): A red/far-red light-responsive bi-stable toggle switch to control gene expression in mammalian cells.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 576
  Illegal BglII site found at 900
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]