Part:BBa_K2715035
Synthetic guide RNA targeting sequence 4 for tcdA promoter
Usage and Biology
This is a 20bp targeting region of a synthetic guide RNA designed to work with the Cas9 protein from S. pyrogenes. It is specific to a portion of the promoter region controlling expression of the Toxin A gene in C. difficile. In our project we aimed to use a nucleolytically inactive Cas9 protein (dCas9) in combination with this unique guide RNA sequence to block the transcription machinery from being able to transcribe tcdA in C. difficile, thus reducing its toxicity.
Multiple guide sequences were designed so that their relative targeting efficiencies could be compared in an assay. The complete list of guides designed can be found below.
Characterisation
The composite part designed to express this synthetic guide RNA can be found here BBa_K2715039, and when expressed in conjugation with a dCas9, will theoretically reduce toxin expression in C. difficile. This part was characterised using an assay for the reporter GusA in place of the C. difficile toxin A. 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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