Protein for regulating lipid metabolism and application thereof
A technology of lipid metabolism and protein, which is applied in the field of protein regulating lipid metabolism and its application, and can solve problems such as unclear functions
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[0107] There is no particular limitation on the preparation method of small interfering RNA, including but not limited to: chemical synthesis method, in vitro transcription method and so on. After knowing the correlation between Patt1 and liver fat metabolism, the small interfering RNA can be prepared to improve liver fat metabolism.
[0108] In addition, the sense and antisense strands contained in the small interfering RNA can be prepared by one or more expression cassettes encoding the sense and antisense strands. When the sense and antisense strands are encoded by a single expression cassette, they can be cleaved from the resulting transcript to form separate sense and antisense strands, which subsequently hybridize to generate double-stranded siRNA.
[0109] The small interfering RNA can be delivered into cells by using appropriate transfection reagents, or can also be delivered into cells by various techniques known in the art.
[0110] The present invention also provid...
Embodiment 1
[0166] Example 1, Construction of Liver-specific Patt1 Knockout Mice
[0167] First, the inventors constructed a liver-specific Patt1 knockout mouse (Patt1 LKO) through the Cre-loxP system. Western results showed that Patt1 protein levels were significantly reduced in the liver of Patt1LKO mice compared with littermate controls (eg figure 1 shown in A); muscle ( figure 1 B), white adipose tissue ( figure 1 C), brain ( figure 1 D), pancreas, kidney and brown adipose tissue ( Figure 7 ) in Patt1 protein levels were not significantly changed compared to littermate controls. This demonstrates the successful construction of Patt1 liver-specific knockout mice.
Embodiment 2
[0168] Example 2. Deletion of Patt1 in the liver mainly caused a significant reduction in fat content
[0169] Long-term observation of body weight and food intake of knockout mice revealed that at 20 weeks of age, male Patt1LKO mice had significantly reduced fat weight and fat content compared with littermate controls (e.g. figure 2 A, 2B shown). Even as early as 6 weeks of age, a significant reduction in body fat content in male Patt1LKO mice could be observed ( Figure 8 ). Lean body mass and lean body mass content of male Patt1LKO mice were somewhat increased compared to littermate controls (e.g. figure 2 Shown in C, 2D), body weight was slightly increased compared with littermate control (two-way ANOVA, p=0.0413) ( figure 2 E). Between 15 and 17 weeks, the food intake of male Patt1LKO mice was slightly increased compared with littermates ( figure 2 F). Through glucose tolerance test and insulin tolerance test, the inventors found that at the age of 15 weeks, the...
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