Bacterial Cell 3-8 Decoder and Cell Computer
A technology of bacterial cells and decoders, applied in the field of synthetic biology, can solve the problem that there is no decoder yet
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Embodiment 1
[0056] This embodiment is used to decode the input conditions of IPTG (first signal), aTc (second signal), and Arabinose (third signal); green fluorescence is used as an indicator signal; CRISPR / dCas9 system is used to realize the indicator Signal inhibition, dCas9 protein is a protein that loses Cas9 cutting activity, cannot cut DNA, but can bind to DNA to prevent transcription, use sgRNA to guide dCas9 protein to bind to the corresponding DNA fragment, thereby preventing transcription from occurring and making gene elements Unable to express; using the flipping function of the DNA site-directed recombinase Cre / loxP system to realize promoter de-repression / repression; using the flipping function of the DNA site-directed recombinase FLP / frt system to realize indication signal de-repression / repression.
[0057] Promoters and signature signals are selected as follows:
[0058] The first promoter: the promoter P activated by the first signal IPTG lac
[0059] Second promoter: p...
Embodiment 2
[0083] In this example, the CRISPR / dCas9 system in Example 1 was replaced with CRISPR / ddCpf1 to achieve the effect of inhibiting gene expression. The ddCpf1 protein is similar to the dCas9 protein in that it cannot cut DNA, but it can bind to DNA to prevent the occurrence of transcription. The sgRNA is used to guide the ddCpf1 protein to bind to the corresponding DNA fragment, thereby preventing the occurrence of transcription and preventing the expression of genetic elements. In this example, the sgRNA is designed according to the requirements of the CRISPR / ddCpf1 system, and E. coli strains or other strains that can ensure the work of CRISPR / ddCpf1 should be selected or constructed. Others are the same as in Example 1.
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