Difference between revisions of "Part:BBa K1150027"
 
(2 intermediate revisions by the same user not shown)
Line 24: Line 24:
 
==Usage and Biology==
 
==Usage and Biology==
  
−
Fusion protein of [https://parts.igem.org/Part:BBa_K1150004 PhyB] and [https://parts.igem.org/Part:BBa_K1150002 KRAB]. This fusion protein PhyB-KRAB is an interaction partner of [https://parts.igem.org/Part:BBa_K1150025 dCas9-PIF]. When crRNA and tracrRNA bind to Cas9, the protein is able to bind complementary DNA. When the system is exposed to red light (660 nm), the Phytochrome B receptor binds to the Phytochrome interaction actor (PIF). So it recruits the dCas9-KRAB protein. The PhyB-PIF binding can be abolished by illumination with far-red light (740nm.) This system enables repression of gene expression induced by red light. Have a look at our [http://2013.igem.org/Team:Freiburg/Project/induction#light light project page].
+
Fusion protein of [https://parts.igem.org/Part:BBa_K1150004 PhyB] and [https://parts.igem.org/Part:BBa_K1150002 KRAB].  
 +
This fusion protein PhyB-KRAB is an interaction partner of [https://parts.igem.org/Part:BBa_K1150025 dCas9-PIF]. When crRNA and tracrRNA bind to [https://parts.igem.org/Part:BBa_K1150000 dCas9], the protein is able to bind complementary DNA. When the system is exposed to red light (660 nm), the Phytochrome B receptor binds to the Phytochrome interaction actor (PIF). So this protein is being recruited by [https://parts.igem.org/Part:BBa_K1150025 dCas9-PIF]. The PhyB-PIF binding can be abolished by illumination with far-red light (740nm).
 +
 
 +
This system enables activation of gene expression induced by red light. For more information, have a look at our [http://2013.igem.org/Team:Freiburg/Project/induction#light light] and [http://2013.igem.org/Team:Freiburg/Project/effector#repression repression] project page.
  
  
  
 
<span class='h3bb'>
 
<span class='h3bb'>
 +
 
==Sequence and Features==
 
==Sequence and Features==
 
</span>
 
</span>

Latest revision as of 20:40, 4 October 2013

uniCAS Red Light Switch Part II - Repressor

CMV:PhyB-Linker-KRAB-NLS:BGH
Function Repression domain of red light

induced gene expression control

Use in Mammalian cells
RFC standard RFC 25
Backbone pSB1C3
Submitted by [http://2013.igem.org/Team:Freiburg Freiburg 2013]

Usage and Biology

Fusion protein of PhyB and KRAB. This fusion protein PhyB-KRAB is an interaction partner of dCas9-PIF. When crRNA and tracrRNA bind to dCas9, the protein is able to bind complementary DNA. When the system is exposed to red light (660 nm), the Phytochrome B receptor binds to the Phytochrome interaction actor (PIF). So this protein is being recruited by dCas9-PIF. The PhyB-PIF binding can be abolished by illumination with far-red light (740nm).

This system enables activation of gene expression induced by red light. For more information, have a look at our [http://2013.igem.org/Team:Freiburg/Project/induction#light light] and [http://2013.igem.org/Team:Freiburg/Project/effector#repression repression] 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 1076
    Illegal BamHI site found at 1158
    Illegal XhoI site found at 1109
    Illegal XhoI site found at 1128
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2803
    Illegal SapI site found at 1325
    Illegal SapI.rc site found at 2767

References

Müller, K. et al. (2013). A red/far-red light-responsive bi-stable toggle switch to control gene expression in mammalian cells. Nucleid Acid Research
Witzgall, R. et al. (1994). The Krüppel-associated box-A domain of zinc finger proteins mediates transcriptional repression. Proc Nati Acad Sci