An animal model for studying atherosclerotic lesions
A technique of atherosclerosis, animal model, applied in aspects, a specific aspect, mice, research of animal models of atherosclerosis, drug, prevention and/or treatment of atherosclerosis, screening for treatment of atherosclerosis Like sclerosing drugs, rodents, / pathological differences in the field, able to address issues such as animal models that do not preserve SPT function
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Embodiment 3
[0094] In yet another embodiment, the present invention provides an animal model for studying metabolic syndrome or insulin resistance, obesity, and diabetes, wherein the genome of the model animal contains a subunit encoding serine palmitoyl-CoA transferase (SPT) Heterozygous division of at least one endogenous gene.
[0095]The invention is further illustrated by the following non-limiting examples.
Embodiment 1
[0097] Preparation of animal models for studying atherosclerosis
[0098] Construction of Sptlc1 Gene Replacement Vector
[0099] The 12kb mouse genome DNA fragment containing Sptlcl exons 7-10 from the mouse 129λ genome library was used for the construction of the targeting vector (Figure 1). PacI-linearized targeting vector was used for electroporation, and G418 was used to selectively select embryonic stem cells (ES). Southern blot analysis and PCR were used to screen targeted ES cells. Genomic DNA was digested with ECoRV, and a 350-bp DNA fragment only 3'to the targeting vector (Figure 2) was used as a probe for Southern blotting.
[0100] The wild type (WT) contains a 7.2 kb fragment, while the recombinant contains a 5.5 kb fragment without exons 7 or 8 ( Figure 1B ). Use primer pair SrSA5 and Neo2 to do PCR. The primer SrSA5 is located outside the short arm and has the sequence 5'-TCAGAGATTCTCCATTGCCACTG-3' (SEQ ID NO: 1). The primer Neo2 is located in the 5'-promoter reg...
Embodiment 2
[0128] Sptlc1 and Sptlc2 lack reduced liver Sptlc1 and Sptlc2 mRNA, quality and activity levels
[0129] Replace exons 7 and 8 with neo gene ( Figure 1A ), using forward selection to target the mouse Sptlcl gene. To screen for homologous elements, the genomic DNA of ES cells was digested with EcoRV. Use the 350bp fragment within intron 6 and outside the target sequence to analyze the Southern blot ( Figure 1B ), 5 out of 150 ES cell clones showed homologous binding. Adding the 5.5-kb signal to the endogenous 7.2-kb signal showed specific binding at the Sptlc1 gene locus ( Figure 1B ). The correctly targeted cells are injected into the C57BL / 6J host blastocyst. Six chimeras (3 males, 3 females) were produced, all of these male chimeras spread the split Sptlcl alleles through the germline. Heterozygous mice obtained by crossing. After screening 300 offspring, no homozygous animals were found. The embryos from 15 to 20 days were screened, and no homozygous mice were found. As expe...
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