MKR mouse model
MKR mouse model was generated and characterized by LeRoith and colleagues. By introducing a point mutation of lysine to arginine, the researchers created dominant negative mutant of human IGF1 receptor. When expressed specifically in skeletal muscle, the transgenic mouse displays severe diabetes due to defects in both insulin and IGF1 signaling. This mouse model is widely used in diabetes research.
Ref: Fernandez AM, et al. Genes & Dev 2001.
MKR 5:
CCG CAA CCT GAC CAT CAA AG
MKR 3:
GCC AGG TTA TGA TGA TGC GA
Product: 221 bp.
Recommended condition:
94C 5 min
94C 30 sec
56C 30 sec
72C 30 sec
X 35 cycles
72C 5 min
4C -
Recommended reaction:
18.5 ul H2O
2.5 ul PCR buffer (10X)
0.5 ul dNTP (10 mM each)
0.5 ul MKR 5 (10 uM)
0.5 ul MKR 3 (10 uM)
0.5 ul Taq (5 U/ul)
2 ul DNA
--------
25 ul
These primers were designed and tested by Wang. These are not the primers shown in the reference publication. These primers have been tested in mouse tail DNA preparations to genotype MKR transgenic animals. These primers only pick up the transgene, human IGF1R, without crossing reaction with endogenous mouse IGF1R. These primers have also been used for quantitative qPCR with cDNA libraries.
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SIRT6 links glucose and lipid metabolism
SIRT1, a histone deacetylase, has drawn considerable attention due to its important roles in metabolic regulation and longevity. SIRT1 is induced by fasting and suppressed by feeding. SIRT1 increases glucose production and beta oxidation in liver to meet energy needs during food deprivation. Another member of SIRT family, SIRT6 follows similar expression patterns as SIRT1. The function of SIRT6 in metabolic regulation remains unknown.
Kim and colleagues found SIRT1 can regulate SIRT6 expression in liver. Specifically, SIRT1 functions together with FOXO3a and stimulates SIRT6 transcription during fasting. The activation of SIRT6 directly suppresses gene expression of metabolic enzymes involved in triglyceride synthesis and glycolysis in liver. Deficiency of SIRT6 causes increases of glucose utilization, reduced beta oxidation and as a result, fatty liver. More importantly, Kim et al found the expression of SIRT6 is decreased in human fatty liver samples which indicates SIRT6 may play critical roles in liver steatosis in clinical settings. This study landed in Cell Metabolism of this month.
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