Difference between revisions of "Part:BBa K1150027"
 
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<partinfo>BBa_K1150027 short</partinfo>
 
<partinfo>BBa_K1150027 short</partinfo>
  
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{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right"
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! colspan="2" style="background:#FFBF00;"|CMV:PhyB-Linker-KRAB-NLS:BGH
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|-
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|'''Function'''
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|Repression domain of red light
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induced gene expression control
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|-
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|'''Use in'''
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|Mammalian cells
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|-
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|'''RFC standard'''
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|[https://parts.igem.org/Help:Assembly_standard_25 RFC 25]
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|-
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|'''Backbone'''
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|pSB1C3<br>
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|-
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|'''Submitted by'''
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|[http://2013.igem.org/Team:Freiburg Freiburg 2013]
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|}
  
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<!-- Add more about the biology of this part here
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==Usage and Biology==
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===Usage and Biology===
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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.
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Fusion protein of [https://parts.igem.org/Part:BBa_K1150004 PhyB] and [https://parts.igem.org/Part:BBa_K1150002 KRAB].  
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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).  
  
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===References===
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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.
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Müller, 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.
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<!-- -->
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<span class='h3bb'>Sequence and Features</span>
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<span class='h3bb'>
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==Sequence and Features==
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</span>
 
<partinfo>BBa_K1150027 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1150027 SequenceAndFeatures</partinfo>
  
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==References==
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<small>
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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 <br>
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Witzgall, R. et al. (1994). The Krüppel-associated box-A domain of zinc finger proteins mediates transcriptional repression. Proc Nati Acad Sci
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</small>
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

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