Regulatory

Part:BBa_K3506021

Designed by: Jingzi Wang,Xinyao Pan   Group: iGEM20_BNU-China   (2020-10-21)
Revision as of 21:44, 26 October 2020 by Yanran (Talk | contribs)


U6 promoter

U6 promoter is used to drive the expression of homing guide RNA(hgRNA) in lineage tracing for eukaryotic systems. We used it to initiate the transcription of hgRNA(sgRNA). The U6 promoter has a clear transcription start point G. The 5′-TTTTT-3′ sequence located downstream of the promoter is used as a transcription termination signal, and the transcription product will be added at the 3′ end of 2 -4 oligomeric U.


Biology and Usage

RNA polymerase III promoters uniquely transcribes small non-coding RNAs, including 5S rRNA , tRNAs, and other essential RNAs such as the U6 snRNA[1].U6 promoter is RNA polymerase III promoters and is a common part for eukaryotic expression systems. We use it to initiate the transcription of hgRNA used in CRISPR/Cas9 sysytem.You also can use the Pol Ⅲ promoter U6 to transcribe short hairpin RNA (shRNA,). The shRNA is cut by Dicer to become siRNA, which can interfere with the expression of target genes through the RNAi system.


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]


References

[1]Wang, Y., Wei, D., Zhu, X. et al. A ‘suicide’ CRISPR-Cas9 system to promote gene deletion and restoration by electroporation in Cryptococcus neoformans. Sci Rep 6, 31145 (2016). https://doi.org/10.1038/srep31145 [2]Li Q, Wang W, Zhao N, Li P, Xin Y, Hu W. Identification and validation of a Schistosoma japonicum U6 promoter. Parasit Vectors. 2017;10(1):281. Published 2017 Jun 5. doi:10.1186/s13071-017-2207-4

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