Mouse model construction method of eGFP-labeled GPR120 positive cells

Abstract

Provided is a mouse model construction method of eGFP-labeled GPR120 positive cells. A guideRNA target sequence aiming at a target locus genome is designed, and in vitro transcription is carried out to obtain guideRNA aiming at the genome; donor DNA recombinant plasmid is constructed; the guideRNA and the donor DNA recombinant plasmid are subjected to oosperm injection to obtain F0-generation mice, and the genotype of the F0-generation mice is identified to obtain positive F0-generation mice achieving correct homologous recombination; the F0-generation mice and wild type mice are copulated to obtain F1-generation mice, and the genotype of the F1-generation mice is identified to obtain positive F1-generation mice achieving correct homologous recombination; positive transgenic mice in the generation F1 are subjected to sib mating to obtain F2-generation mice, and homozygous transgenic mice are obtained through screening and identifying. According to the method, GPR120 expression cell express eGFP at the same time, and therefore GPR120 positive cells are distinguished under laser excitation.

Description

technical field [0001] The invention belongs to the field of transgenic technology, and in particular relates to a method for constructing a mouse model of eGFP-marked GPR120-positive cells. Background technique [0002] GPR120 (G-protein coupled receptor 120; also known as free fatty acid receptor 4, FFAR4) is a member of the fatty acid receptor family and belongs to G protein coupled receptors (GPCR), which can be activated by long-chain fatty acids, especially n-3 Unsaturated fatty acids have the strongest activation ability. GPR120 is expressed in many tissues such as the brain, pituitary, lung, tongue, gastrointestinal tract, and adipose tissue, and is mainly distributed in various endocrine cells, adipocytes, macrophages, osteoblasts and osteoclasts, and taste bud cells in the gastrointestinal tract. GPR120 activation can stimulate the secretion of gastrointestinal hormones such as GLP-1, CCK, and GIP. In 2010, a research team in the United States found that GPR120 c...

AI Technical Summary

Problems solved by technology

Currently, methods for labeling and separating live cells include flow cytometric antibody labeling and plasmid transfection labeling. The main problems of these methods are false positive labeling, high antibody consumption costs, large operational differences between groups, and large changes in the success rate of labeling.

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