Difference between revisions of "Part:BBa K1150027"
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===References===
 
===References===
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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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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
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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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Revision as of 12:30, 2 October 2013

uniCAS Red Light Switch Part II - Repressor

uniCAS red Light Switch Part II - Repressor
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 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.

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

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