Difference between revisions of "Part:BBa K1150025"
Line 3: Line 3:
  
 
{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right"
 
{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right"
−
! colspan="2" style="background:#FFBF00;"|CMV:dCas9-PIF6:BGH
+
! colspan="2" style="background:#FFBF00;"|CMV:HA-NLS-dCas9-Linker-PIF6-NLS:BGH
 
|-
 
|-
 
|'''Function'''
 
|'''Function'''

Revision as of 12:57, 4 October 2013

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 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 dCas9-version is unable to cleave DNA. PIF6 is human codon-optimized. Have a look at our [http://2013.igem.org/Team:Freiburg/Project/induction#light light project page].



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.