Difference between revisions of "Part:BBa K1150017"

 
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__NOTOC__
 
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{| style="color:black" cellpadding="6" cellspacing="1" border="2" align="right"
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! colspan="2" style="background:#FFBF00;"|pSV40:HA-NLS-dCas9-G9a-NLS:tBGH
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|-
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|'''Function'''
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|DNA binding protein
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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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|'''Organism'''
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|<i>Streptococcus pyogenes</i>
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|-
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|'''Source'''
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|Feng Zhang, Addgene<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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<partinfo>BBa_K1150017 short</partinfo>
 
<partinfo>BBa_K1150017 short</partinfo>
  
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Cas9 is a protein that binds DNA via an protein-RNA-DNA interaction. This version of Cas9 is working as a carrier for effectors that want to be brought to DNA in a sequence specific manner. The native nikase activity of the protein has been mutated so that it allows DNA binding without cutting. The sequence is human codon optimized.
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The protein is part of team Freiburg's (2013) the uniCAS toolkit.
  
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<!-- Add more about the biology of this part here
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The usage of the strong [https://parts.igem.org/Part:BBa_K1150015 CMV promoter] enables this device to be expressed in a strong manner. If weaker expression levels are needed, we recommend using our uniCAS Histone Modificator device with an [https://parts.igem.org/Part:BBa_K1150018 SV40 promoter].
 
===Usage and Biology===
 
===Usage and Biology===
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The dCAS9 protein is able to be targeted to several loci at once as it interacts with small RNAs to build up a complex that will interact with complementary DNA strands. Its origin is the adaptive immune system of <i>Streptococcus pyogenes </i> called CRISPR. Hijacking this system leads to a whole new approach for multiple gene targeting.
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The iGEM team Freiburg 2013 combined effectors with this protein to create a toolkit, that is able to activate or repress specific and multiple target loci.
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This approach offers new possibilities for fundamental epigenetic research, tissue engineering and cancer research.
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1150017 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K1150023 SequenceAndFeatures</partinfo>
  
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  
 
===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K1150017 parameters</partinfo>
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<partinfo>BBa_K1150023 parameters</partinfo>
 
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Revision as of 09:49, 2 October 2013



pSV40:HA-NLS-dCas9-G9a-NLS:tBGH
Function DNA binding protein
Use in Mammalian cells
RFC standard RFC 25
Backbone pSB1C3
Organism Streptococcus pyogenes
Source Feng Zhang, Addgene
Submitted by [http://2013.igem.org/Team:Freiburg Freiburg 2013]

dCas9 with CMV promoter

Cas9 is a protein that binds DNA via an protein-RNA-DNA interaction. This version of Cas9 is working as a carrier for effectors that want to be brought to DNA in a sequence specific manner. The native nikase activity of the protein has been mutated so that it allows DNA binding without cutting. The sequence is human codon optimized. The protein is part of team Freiburg's (2013) the uniCAS toolkit.

The usage of the strong CMV promoter enables this device to be expressed in a strong manner. If weaker expression levels are needed, we recommend using our uniCAS Histone Modificator device with an SV40 promoter.

Usage and Biology

The dCAS9 protein is able to be targeted to several loci at once as it interacts with small RNAs to build up a complex that will interact with complementary DNA strands. Its origin is the adaptive immune system of Streptococcus pyogenes called CRISPR. Hijacking this system leads to a whole new approach for multiple gene targeting.

The iGEM team Freiburg 2013 combined effectors with this protein to create a toolkit, that is able to activate or repress specific and multiple target loci.

This approach offers new possibilities for fundamental epigenetic research, tissue engineering and cancer research.


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 664
    Illegal BglII site found at 5139
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]