Difference between revisions of "Part:BBa K3506021"

Revision as of 17:33, 27 October 2020

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 initiate 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 recognized and digested by Dicers to generate siRNA, which can interfere with the expression of target genes through the RNAi system.

Sequence and Features

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).