Difference between revisions of "Part:BBa K2217024:Design"

 
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===Design Notes===
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==Design Notes==
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Composite Part
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Part BBa_K2217019 was composed of CMV Enhancer as well as CMV Promoter (BBa_K2217000 and  BBa_K2217006 respectively) as constitutive promoters to enhance the transcription process of the Cas9 (BBa_K1218011) part which was transcribed using  T7 Promoter (BBa_K2217007) following Protocol[1]. The design aimed at generating Homology Directed Repair instead of Non-Homologous end Joining repair of Cas9 by knocking in the circular RNA as a Competing endogenous RNA enhancing its transcription to regulate miRNA action, so we designed Homology repair template (BBa_K2217009) to be transfected on a separate donor vector.
  
  
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Synthesized Fragments by IDT
 
Synthesized Fragments by IDT
  
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===References===
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==References==
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[1] Romanienko PJ, Giacalone J, Ingenito J, et al. A Vector with a Single Promoter for In Vitro Transcription and Mammalian Cell Expression of CRISPR gRNAs. Fujii H, ed. PLoS ONE. 2016;11(2):e014836.

Revision as of 16:10, 23 October 2017


CMV Enhancer + CMV Promoter


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]


Design Notes

Part BBa_K2217019 was composed of CMV Enhancer as well as CMV Promoter (BBa_K2217000 and BBa_K2217006 respectively) as constitutive promoters to enhance the transcription process of the Cas9 (BBa_K1218011) part which was transcribed using T7 Promoter (BBa_K2217007) following Protocol[1]. The design aimed at generating Homology Directed Repair instead of Non-Homologous end Joining repair of Cas9 by knocking in the circular RNA as a Competing endogenous RNA enhancing its transcription to regulate miRNA action, so we designed Homology repair template (BBa_K2217009) to be transfected on a separate donor vector.


Source

Synthesized Fragments by IDT

References

[1] Romanienko PJ, Giacalone J, Ingenito J, et al. A Vector with a Single Promoter for In Vitro Transcription and Mammalian Cell Expression of CRISPR gRNAs. Fujii H, ed. PLoS ONE. 2016;11(2):e014836.