(Pro) resin receptor (P) RR gene and application of inhibitor of gene
A technology of gene and gene expression, which is applied in the determination/testing of microorganisms, medical preparations containing active ingredients, metabolic diseases, etc., and can solve problems such as unclear effects
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Embodiment 1
[0052] 1. Obtain control group mice and experimental group mice and carry out genotype identification
[0053] As experimental mice, C57 mice were used. Alb-cre + / + Male mice were purchased from Nanjing model animals, and the mice specifically expressed Cre recombinase in the liver. (P)RR Flox / WT Female mice were purchased from Nanjing model animals. The mice were transgenic mice with loxP sites at both ends of exon 2 of the (P)RR gene, wherein exon 2 of the (P)RR gene together with its two ends The loxP site is referred to as the Flox fragment or (P)RR flox in the present invention. (P)RR Flox / WT The mouse preparation process includes constructing an expression vector containing (P)RR flox, transfecting mouse embryonic stem cells, performing homologous recombination, screening out cells expressing (P)RR flox, and microinjecting the cells into early stage mice Embryos were transplanted into pseudopregnant mice, and transgenic mice expressing (P)RR flox were obtained throu...
Embodiment 2
[0086] 1. Inhibition of (P)RR expression by tail vein injection of (P)RR liver-specific antisense oligonucleotides (ie, knockdown)
[0087] Common C57 mice were used as the experimental objects. The mice in the blank control group were injected with PBS through the tail vein, the mice in the scrambled control group were injected with the liver-specific scrambled sequence, and the mice in the experimental group were injected with (P)RR liver-specific reaction through the tail vein. The sense oligonucleotide drug was injected once a week, and the injection amount of both the control group and the experimental group was 3 mg / kg / week.
[0088] Wherein, the liver-specific scrambled sequence is: 5'-GGCCAATACGCCGTCA-3'; the sequence of (P)RR liver-specific antisense oligonucleotide is: 5'-AGATATTGGTCCATTT-3'.
[0089] 2. Study the function of the liver (P)RR gene through the obese mouse model induced by high-fat diet.
[0090] The mice in the three groups were all male and 8 weeks o...
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