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CRISPRi/a
CRISPRi: bcl-2 repression
CRISPRi is to use dCas9-KRAB to block the RNA polymerase transcription elongation and initialization. We till several sgRNAs or single sgRNA in the bcl-2 promoter to obtaion optimal repression. We create the CRISPRi model with mass action laws in order to gain fold repression of bcl-2
CRISPRa: bax activation
CRISPRa is to use dCas9-VP64 to activate the transcription initialization. We use the similar stragegy to obtain optimal activation. And we create CRISPRa model with mass action laws in order to gain fold activation of bax.
The tumor specific promoter in our project is with an ampilification system. It contains a minimal promoter targeted by three dCas9-VP64-sgRNA complex. We build a two-step-transcription-activation model to analyze whether it can promote enough dCas9-VP64.
We build up a model for tumor specific promoter that expresses the CRISPR/dCas9-VP96 system.
The dCas9-VP64-GFP expression rate under EF1α promoter, survivin promoter, and Bax_1 promoter.