Difference between revisions of "Part:BBa K1150031"
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Revision as of 12:17, 4 October 2013

uniCAS UV Light Switch Part II - Repressor

CMV:COP1-KRAB:BGH
Function Repression domain of UVB light

induced gene expression control

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

The fusion protein COP1-KRAB is an interaction partner of dCas9-UVR8 and part of the UVB light controlled mechanism repress gene expression. First the dCas9-UVR8 fusion protein binds specifically to a DNA target sequence. If the system is exposed to UVB light (311 nm), the UVR8 receptor of UV changes its configuration and recruits this COP1-KRAB protein. This represses gene expression controlled by UVB light.

Biology and Usage

E3-ubiquitin ligase Constitutively Photomorphogenic 1 (COP1) is the interaction partner of ultraviolet B (UVB) light receptor UVR8 [1]. When UVB light is perceived by the UVR8 photoreceptor it monomerizes and therefore changes in its open confuguration. Not till then it recruits COP1. For this UVB light controled gene activation system you need a combination of these BioBricks: Cas9-UVR8, COP1-KRAB and RNaimer.

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
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1373
    Illegal BsaI.rc site found at 1450
    Illegal BsaI.rc site found at 1876
    Illegal SapI.rc site found at 962
    Illegal SapI.rc site found at 1840

References

[1] Rizzini L., Favory J.-J., Cloix C., Faggionato D., O'Hara A., Kaiserli E., Baumeister R., Schäfer E., Nagy F., Jenkins G. I., Ulm R. (2011). Perception of UV-B by the Arabidopsis UVR8 protein. Science 332