Difference between revisions of "Part:BBa K1150025"
 
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{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right"
 
{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right"
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! colspan="2" style="background:#FFBF00;"|uniCAS UV Light Switch Part II - Activator
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! colspan="2" style="background:#FFBF00;"|CMV:HA-NLS-dCas9-Linker-PIF6-NLS:BGH
 
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|'''Function'''
 
|'''Function'''
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|Activation domain of UVB light
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|dCas9-protein linked to PIF6
 
induced gene expression control  
 
induced gene expression control  
 
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=Introduction=
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==Introduction==
  
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Our construct contains the standardized version of Cas9 linked to PIF6. This device is one of two parts that you need to enable red-light induced gene targeting. Cotransfection of a PhyB+effector device, and target RNA allows you to recruit any desired effector to your desired Gene locus. Illumination with red light (660nm) will start the interaction and illumination with far red light (740nm) will stop the interaction.
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Our construct contains the standardized version of [https://parts.igem.org/Part:BBa_K1150000 dCas9] linked to [https://parts.igem.org/Part:BBa_K1150005 PIF6]. This device is one of two parts that you need to enable red-light induced gene targeting. Cotransfection of a PhyB+effector ([https://parts.igem.org/Part:BBa_K1150026 activation], [https://parts.igem.org/Part:BBa_K1150027 repression] or [https://parts.igem.org/Part:BBa_K1150028 epigenetical repression]) device and target RNA makes it possilbe to recruit any desired effector to your desired gene locus. Illumination with red light (660nm) will start the interaction and illumination with far red light (740nm) will stop the interaction.
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The Cas9-version used here is human codon optimized and the mutated version, unable to cleave DNA. PIF6 is human codon-ptimized.
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The here used, mutated Cas9-version is human codon optimized and unable to cleave DNA. PIF6 is human codon-optimized, too. For more information and experimental data have a look at our [http://2013.igem.org/Team:Freiburg/Project/induction#light light project page].
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=References:=
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Muller, 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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<!-- Add more about the biology of this part here
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==Data==
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===Usage and Biology===
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Data for this part are shown on the page of [https://parts.igem.org/Part:BBa_K1150026 Part II]
  
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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_K1150025 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1150025 SequenceAndFeatures</partinfo>
  
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==References:==
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Muller, 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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<!-- Add more about the biology of this part here
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==Usage and Biology==
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Latest revision as of 01:53, 5 October 2013

uniCAS Red Light Switch Part I - Stimulator

CMV:HA-NLS-dCas9-Linker-PIF6-NLS:BGH
Function dCas9-protein linked to PIF6

induced gene expression control

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

Introduction

Our construct contains the standardized version of dCas9 linked to PIF6. This device is one of two parts that you need to enable red-light induced gene targeting. Cotransfection of a PhyB+effector (activation, repression or epigenetical repression) device and target RNA makes it possilbe to recruit any desired effector to your desired gene locus. Illumination with red light (660nm) will start the interaction and illumination with far red light (740nm) will stop the interaction. The here used, mutated Cas9-version is human codon optimized and unable to cleave DNA. PIF6 is human codon-optimized, too. For more information and experimental data have a look at our [http://2013.igem.org/Team:Freiburg/Project/induction#light light project page].

Data

Data for this part are shown on the page of Part II


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 900
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

References:

Muller, 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.