A method of constructing a cell model used for detecting a heat source, a cell model and a heat source detection kit

摘要

The present invention provides a method for constructing a cell model used for detecting a heat source, a cell model, and a heat source detection kit. The cell model induces the formation of double bond disruption at a specific site of the genome using CRISPR / CAS9. Using the principle of homologous recombination repair, TLR4 and CD14-MD2 are inserted at designated positions of two chromosomes in the cell line We followed a fluorescently-traced cell model in which the final successfully constructed TLR4 / CD14 / MD2 was inserted at the designated position by green fluorescence GFP and red fluorescence RFP, respectively, and the LPS-stimulated cell model was ELISA, Western The release of IL-6, TNF-a cell factor can be detected by Western blot, mass spectrometry and magnetic bead method. The cell model has good stability and sensitivity, and the lowest detection limit is 0.005 EU / mL , Which is much lower than 0.025 EU / mL of the pellet reagent method.;