Far-red light regulation and control gene expression loop control system as well as building method and application thereof
A technology of gene regulation and loop control, applied in multidisciplinary fields, can solve problems such as hypoglycemia, and achieve the effect of low expression background, high regulation expression multiple, and great application value
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Embodiment 1
[0072] Example 1, Construction of Gene Circuit Regulatory System Components for Regulating Transgene Expression with Far-red Light
[0073] This example includes the construction method of representative elements in the gene loop remote regulation system for regulating the expression of transgenes by far-red light, but it does not limit the protection scope of the present invention. The detailed design scheme and steps are shown in Table 1.
Embodiment 2
[0074] Example 2. Far-red light regulates transgene expression. Far-red light regulates gene expression loop control system. Photoreceptors expressed by different promoters
[0075] The first step, plasmid construction. The details of the plasmid construction in this example are shown in Table 1. The second step is to inoculate the cells. The third step is transfection. Within 16 to 24 hours of inoculating the cells, divide two 24-well plates into a dark group and a light group, and each group is equally divided into 1-5 groups. Within 16 to 24 hours of inoculating cells, 0.1 μg pWS50 (expressed by the promoter SV40) was added to the first group of the dark group and the light group, 0.1 μg pWS189 (expressed by the promoter CMV) was added to the second group, and 0.1 μg pWS189 (expressed by the promoter CMV) was added to the third group. 0.1 μg pWS51 (expressed by the promoter hEF1α) in the middle group, 0.1 μg pWS55 (expressed by the promoter mPGK) in the 4th panel, 0.1 μg...
Embodiment 3
[0077] Example 3, far-red light regulates the gene loop control system of transgene expression Different amounts of processors
[0078] The first step, plasmid construction. The details of the plasmid construction in this example are shown in Table 1. The second step is to inoculate the cells. The third step is transfection. Within 16 to 24 hours of inoculating the cells, divide two 24-well plates into a dark group and a light group, and each group is equally divided into 1-8 groups. Within 16 to 24 hours of inoculating cells, in the dark group and the light group, 0ng pSTING was added to the first group, 5ng pSTING was added to the second group, 10ng pSTING was added to the third group, 20ng pSTING was added to the fourth group, and 20ng pSTING was added to the fifth group. 40ng pSTING, 60ng pSTING in panel 6, 80ng pSTING in panel 7, 100ng pSTING in panel 8 were mixed with 100ng of far-red photoreceptor pWS189, 100μg effector pWS67, PEI transfection reagent with serum-free...
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