Team:UNOTT/Parts
PARTS:
Our BioBricks
For this year's iGEM competition, our team have submitted a promoter-sgRNA inhibition/expression library as parts, using RFP as the model expression protein. We hope this will allow teams to choose sgRNA/Promoter combinations for use in prokaryotes easier, especially with increased interest in CRISPR/Cas systems. The parts submitted rely upon a deactivated Cas9 nuclease (dCas9) present which uses the gRNA to target complementary seequences using Watson-Crick base-pairing. Whereas Cas9 nuclease cuts the target sequence, dCas9 blocks RNA polymerase by steric hindrance without cutting the DNA strands. This causes repression of the gene.
The above schematic shows the gRNA forming a complex with dCas9
To see data for characterisation of these parts please look at Step 6: CRISPRi on Experiments page.
| Name | Type | Paired RFP BioBrick | Description | Bp |
|---|---|---|---|---|
| BBa_K2284001 | gRNA 1 | P1-sRFP-T2 | The first guide RNA as part of the UNOTT teams' gRNA library to target a respective RFP promoter. | 181 |
| BBa_K2284002 | gRNA 2 | P2-sRFP-T2 | The second guide RNA as part of the UNOTT teams' gRNA library to target a respective RFP promoter. | 181 |
| BBa_K2284003 | gRNA 3 | P3-sRFP-T2 | The third guide RNA as part of the UNOTT teams' gRNA library to target a respective RFP promoter. | 181 | BBa_K2284004 | gRNA 4 | P4-sRFP-T2 | The fourth guide RNA as part of the UNOTT teams' gRNA library to target a respective RFP promoter. | 181 | BBa_K2284005 | gRNA 5 | P5-sRFP-T2 | The fifth guide RNA as part of the UNOTT teams' gRNA library to target a respective RFP promoter. | 181 | BBa_K2284000 | gRNA 0 | PE-sRFP-T2 | The sixth guide RNA as part of the UNOTT teams' gRNA library to target a respective RFP promoter. This gRNA intreracts with an empty promoter. | 181 |
References
1. Nielsen, A. and Voigt, C. (2014). Multi-input CRISPR/Cas genetic circuits that interface host regulatory networks. Molecular Systems Biology, 10(11), pp.763-763.
