Part:BBa_K1150025

uniCAS Red Light Switch Part I - Stimulator

CMV:HA-NLS-dCas9-Linker-PIF6-NLS:BGH
Function	dCas9-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 dCas9 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 (activation, repression or epigenetical repression) device and target RNA makes it possilbe 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 here used, mutated Cas9-version is human codon optimized and unable to cleave DNA. PIF6 is human codon-optimized, too. For more information and experimental data have a look at our [http://2013.igem.org/Team:Freiburg/Project/induction#light light project page].

Data

Data for this part are shown on the page of Part II

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]

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.