Difference between revisions of "Part:BBa K1923007"
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<partinfo>BBa_K1923007 short</partinfo>
 
<partinfo>BBa_K1923007 short</partinfo>
  
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Gal4-dCas9-GFP fusion protein is a fusion enzyme.
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This composite part is a fusion protein composed of GAL4, triple FLAG tag, two SV40 nuclear localization sequence,dCas9 and GFP. Triple FLAG tag and GFP is designed for observation and expression detection. The protein produced by this parts can be sequestered in the cytoplasm with the help of sgRNA and PAMmer, utilizing dCas9's mRNA binding ability. However, when there is a mutation in the target sequence of the mRNA, sv40 NLS would drag this protein into the nuclear. Then GAL4 would active the gene downstream UAS promoter. Thus this parts can be used for gene mutation surveillance <sup>[1]</sup>.
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<strong>Reference:</strong>
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<br>
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[1] Nelles D A, Fang M Y, O’Connell M R, et al. Programmable RNA tracking in live cells with CRISPR/Cas9[J]. Cell, 2016, 165(2): 488-496.
  
 
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Revision as of 20:34, 18 October 2016


Gal4BD-NLS-FLAG-EGFP-dCas9-NLS-Gal4AD encoding gene

This composite part is a fusion protein composed of GAL4, triple FLAG tag, two SV40 nuclear localization sequence,dCas9 and GFP. Triple FLAG tag and GFP is designed for observation and expression detection. The protein produced by this parts can be sequestered in the cytoplasm with the help of sgRNA and PAMmer, utilizing dCas9's mRNA binding ability. However, when there is a mutation in the target sequence of the mRNA, sv40 NLS would drag this protein into the nuclear. Then GAL4 would active the gene downstream UAS promoter. Thus this parts can be used for gene mutation surveillance [1].

Reference:
[1] Nelles D A, Fang M Y, O’Connell M R, et al. Programmable RNA tracking in live cells with CRISPR/Cas9[J]. Cell, 2016, 165(2): 488-496.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 2394
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 4673
    Illegal XhoI site found at 220
    Illegal XhoI site found at 567
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 139