Difference between revisions of "Part:BBa K2549030"

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=====It works as we designed =====
 
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[[File:sTF-test.png|none|540px|thumb|'''Flow cytometry results suggest that it can be repressed significantly by ZF42.10-KRAB.''']]
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[[File:sTF-test.png|none|480px|thumb|'''Flow cytometry results of transcriptional repressor interaction with zinc finger-based binding sites. DBD, DNA binding domain which is zinc finger in our assay. SD, silencing-form transcriptional domain. RE, responsive elements. MFI, median fluorescence intensity.''']]
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It is obvious that the three zinc finger-based operators can be significantly repressed by their specific zinc finger-based transcriptional repressor, respectively.
  
  

Revision as of 10:23, 17 October 2018

8*ZF42.10-CMV

This part is one of the response elements of our amplifier, also executing the combiner function. 8*ZF42.10 binding sites (Part:BBa_K2549012) is assembled using two 4*ZF42.10 binding sites (Part:BBa_K2446005) with a biobrick scar between them. CMV (Part:BBa_K2549050) is a promotor which has a high-level constitutive expression. This part can switch off the expression of gene downstream after induced by our zinc finger-based transcription repressor.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Biology

Synthetic promotor operators regulated by artificial zinc finger-based transcription factors

Khalil AS et al have reported several synthetic promotor operators which can interact with artificial zinc finger-based transcription factors with high specificity and high orthogonality[1].

Khalil AS et al stated:sTFs constructed from OPEN-engineered ZFs are orthogonal to one another. sTF43-8 activated noncognate Promoter21-16 due to the fortuitous creation of a sequence that is significantly similar to the binding sequence of 43-8, when the downstream BamHI restriction site is considered.

Characterization

It works as we designed
Flow cytometry results of transcriptional repressor interaction with zinc finger-based binding sites. DBD, DNA binding domain which is zinc finger in our assay. SD, silencing-form transcriptional domain. RE, responsive elements. MFI, median fluorescence intensity.

It is obvious that the three zinc finger-based operators can be significantly repressed by their specific zinc finger-based transcriptional repressor, respectively.


References

  1. ↑ A synthetic biology framework for programming eukaryotic transcription functions. Khalil AS, Lu TK, Bashor CJ, ..., Joung JK, Collins JJ. Cell, 2012 Aug;150(3):647-58 PMID: 22863014; DOI: 10.1016/j.cell.2012.05.045