Difference between revisions of "Part:BBa K1150027"

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

@@ Line 3: / Line 3: @@
 {| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right"
-! colspan="2" style="background:#FFBF00;"|CMV:PhyB-KRAB:BGH
+! colspan="2" style="background:#FFBF00;"|CMV:PhyB-Linker-KRAB-NLS:BGH
 |-
 |'''Function'''
@@ Line 24: / Line 24: @@
 ==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'>
 ==Sequence and Features==
 </span>