CRISPRi device with Ptac expressed gRNA and Anderson strong (J23100)- expressed lacI
- 10COMPATIBLE WITH RFC
- 12Illegal NheI site found at 310
Illegal NheI site found at 333
Illegal NheI site found at 1466
Illegal NheI site found at 4626
Illegal NheI site found at 4649
- 21Illegal BglII site found at 5830
Illegal BamHI site found at 3745
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000COMPATIBLE WITH RFC
After we clarified the correlation between lacI dosage and its induction efficiency, we furthered the experiments by adding CRISPRi to the system. Instead of sfGFP, the IPTG induction devices with different levels of lacI generator is now going to produce a gRNA that targets the promotor of a constitutive sfGFP generator (BBa_K2558206, BBa_K2558207, BBa_K2558208). By adding IPTG, we can induce the transcription of gRNA. Binding with constantly expressed dCas9, gRNA is going to inhibit the expression of sfGFP. We will be able to observe how this process can operate under different levels of lacI expression.
 Gilbert LA et al. Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation. Cell. 2014 Oct 23;159(3):647-61.
 iGEM parts: PHIF repressible promotor (http://parts.igem.org/Part:BBa_K1725000)