Difference between revisions of "Part:BBa K3365000"
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<partinfo>BBa_K3365000 short</partinfo>
 
<partinfo>BBa_K3365000 short</partinfo>
  
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An RNAP &#969; subunit, RpoZ is fused to dCas9 C-terminal.
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An RNA polymerase ω subunit is fused to the C-terminal dCas9.
  
 
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The dCas9 is a Cas9 nuclease mutant. Mutations D10A and H840A in the RucC and HNH domains, respectively, abolish cleavage but do not impair DNA binding. The dCas9 provides a simple and robust technology for gene repression and activation, and can target almost any DNA sequence aided by the sgRNA. The ω subunit can activate transcription by recruiting the RNAP holoenzyme. The fusion between dCas9 and ω subunit can activate gene transcription downstream of the protospacer. The CRISPR interference inhibits transcription by sterically blocking the RNA polymerase(RNAP).
 
The dCas9 is a Cas9 nuclease mutant. Mutations D10A and H840A in the RucC and HNH domains, respectively, abolish cleavage but do not impair DNA binding. The dCas9 provides a simple and robust technology for gene repression and activation, and can target almost any DNA sequence aided by the sgRNA. The ω subunit can activate transcription by recruiting the RNAP holoenzyme. The fusion between dCas9 and ω subunit can activate gene transcription downstream of the protospacer. The CRISPR interference inhibits transcription by sterically blocking the RNA polymerase(RNAP).
  
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[[File:Gene_circuit_of_dCas9_with_subnit_Omega.png]]
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<center>[[File:Gene_circuit_of_dCas9_with_subnit_Omega.png]]</center>
  
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<b>Fig:</b>The gene circuit of this part
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<center><b>Fig:</b>The gene circuit of this part</center>
  
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[[File:Working_mode_of_dCas9_with_subnit_Omega.png]]
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<center>[[File:Working_mode_of_dCas9_with_subnit_Omega.png]]</center>
  
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<b>Fig:</b>The figure above is shown how this part work in bacterial. The geneⅢ mentioned can be changed to any gene sequence depends on designed sgRNA.
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<center><b>Fig:</b>The figure above is shown how this part work in bacterial. The geneⅢ mentioned can be changed to any gene sequence depends on designed sgRNA.</center>
  
 
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Revision as of 17:53, 27 October 2020


dCas9-ω

An RNA polymerase ω subunit is fused to the C-terminal dCas9.

Usage and Biology

The dCas9 is a Cas9 nuclease mutant. Mutations D10A and H840A in the RucC and HNH domains, respectively, abolish cleavage but do not impair DNA binding. The dCas9 provides a simple and robust technology for gene repression and activation, and can target almost any DNA sequence aided by the sgRNA. The ω subunit can activate transcription by recruiting the RNAP holoenzyme. The fusion between dCas9 and ω subunit can activate gene transcription downstream of the protospacer. The CRISPR interference inhibits transcription by sterically blocking the RNA polymerase(RNAP).

Gene circuit of dCas9 with subnit Omega.png
Fig:The gene circuit of this part
Working mode of dCas9 with subnit Omega.png
Fig:The figure above is shown how this part work in bacterial. The geneⅢ mentioned can be changed to any gene sequence depends on designed sgRNA.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1340
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1340
    Illegal NheI site found at 1099
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1340
    Illegal BamHI site found at 3378
    Illegal BamHI site found at 4212
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1340
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1340
  • 1000
    COMPATIBLE WITH RFC[1000]