Regulatory

Part:BBa_K3506021

Designed by: Jingzi Wang,Xinyao Pan   Group: iGEM20_BNU-China   (2020-10-21)
Revision as of 10:17, 27 October 2020 by Panxinyao (Talk | contribs)


U6 promoter

U6 promoter is used to drive the transcription 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 transcription start point G. The 5′-TTTTT-3′ sequence located downstream of the promoter is used as a transcription termination signal, and 2-4 oligomeric U will be added at the 3’ end of the transcription product.


Biology and Usage

RNA polymerase III promoters initiates the transcription of small non-coding RNAs, including 5S rRNA , tRNAs, and other small RNAs such as the U6 snRNA[1]. U6 promoter is recognized by RNA polymerase III. We use it to initiate the transcription of hgRNA in CRISPR/Cas9 system. This promoter can also be used to transcribe short hairpin RNA (shRNA). The shRNA is recongnized and digested by Dicers to generate 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).

[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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