Part:BBa_K2080003
bax-sgRNA3
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, 37 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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