Difference between revisions of "Part:BBa K2381024"
 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K2381024 short</partinfo>
 
<partinfo>BBa_K2381024 short</partinfo>
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<p>
 
<p>
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It’s a gRNA sequence containing a promoter of <partinfo>BBa_J23119</partinfo>, 20bp targeting at <i>oriC</i> and a full-length gRNA scaffold.</p>
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It’s a gRNA sequence containing a promoter of BBa_J23119, 20bp targeting at oriC and a full-length gRNA scaffold.
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[[File:T--HZAU-China--gRNA_function.png|200px|thumb|left|Its function is proved by OD measurement after cotransformed with plasmid containing dCas9.We can observe tight repression on cell growth. ]]
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</p>
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[[File:T--HZAU-China--gRNA_qPCR.png|200px|thumb|left|Further investigation is conducted by qPCR and flow cytometric. Its effect is even better than rapamycin, a well-used chemical molecule for replication inhibition.]]
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<img src="https://static.igem.org/mediawiki/parts/d/d2/T--HZAU-China--gRNA_function.png" width="700px"/>
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</br>
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Its function is proved by OD measurement after cotransformed with plasmid containing dCas9.We can observe tight repression on cell growth.
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</br>
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</br>
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</br>
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<img src="https://static.igem.org/mediawiki/parts/2/26/T--HZAU-China--gRNA_qPCR.png" width="700px"/>
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</br>
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Further investigation is conducted by qPCR and flow cytometric. Its effect is even better than rapamycin, a well-used chemical molecule for replication inhibition.
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</br>
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</br>
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<h3>References</h3>
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1. Wiktor, J., Lesterlin, C., Sherratt, D. J., & Dekker, C. (2016). CRISPR-mediated control of the bacterial initiation of replication. Nucleic Acids Res, 44(8), 3801-3810.
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2. Schmidl, S. R., Sheth, R. U., Wu, A., & Tabor, J. J. (2014). Refactoring and optimization of light-switchable Escherichia coli two-component systems. ACS Synth Biol, 3(11), 820-831.
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Latest revision as of 03:36, 2 November 2017


J23119-gRNA (tightly target at oriC to inhibit cell replication)

It’s a gRNA sequence containing a promoter of BBa_J23119, 20bp targeting at oriC and a full-length gRNA scaffold.


Its function is proved by OD measurement after cotransformed with plasmid containing dCas9.We can observe tight repression on cell growth.



Further investigation is conducted by qPCR and flow cytometric. Its effect is even better than rapamycin, a well-used chemical molecule for replication inhibition.

References

1. Wiktor, J., Lesterlin, C., Sherratt, D. J., & Dekker, C. (2016). CRISPR-mediated control of the bacterial initiation of replication. Nucleic Acids Res, 44(8), 3801-3810. 2. Schmidl, S. R., Sheth, R. U., Wu, A., & Tabor, J. J. (2014). Refactoring and optimization of light-switchable Escherichia coli two-component systems. ACS Synth Biol, 3(11), 820-831.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 47
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]