Device

Part:BBa_K2080002

Designed by: Haojian Li   Group: iGEM16_SCUT-China_B   (2016-09-22)


bax-sgRNA2

This guide RNA is a hundred base-long molecule with a dCas9-VP64 scaffold sequence, which can guide dCas9-VP64 to its complementary part of a double-strand DNA. In our project, this gRNA is targeting to the promoter of bax, a proapoptosis protein, which can activate the transcription of bax.

Usage and Biology

gRNA sequence is complementary to the promoter of bax, 171 bp upstream of the transcription start site.

Characterization

This part was designed for dCas9-VP64 targetting bax promoter then upregulated the transcription level of bax. The upregulation of bax induced apoptosis of A549. The transcription level of bax is detected by real-time PCR(Fig.1).The apoptosis index was detected by Annexin V-FIFC/PI after dCas9-VP64 and gRNA cotransfection after 48 hour(Fig.2)

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]


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Categories
Parameters
None