Conditional knockout of MGAT1 mouse model
Conditional knockout of MGAT1 mouse model was generated and characterized by Stanley and collegues. MGAT1 gene encodes GlcNac transferase I, which initiates the synthesis of complex and bybrid glycans. Loss of MGAT1 causes absence of branch glycans, but does not affect high mannose glycan synthesis and glycoprotein folding & secretion.
Ref: Shi S, et al. Mol Cell Biol 2004.
MGAT 5:
TGC AAG CCA ACA CTT GTC TC
MGAT 3:
GAG ACC TGC TTA CTG CAG CC
Product: Floxed allele of 561 bp and wild type allele of 421 bp.
Recommended condition:
94C 5 min
94C 30 sec
58C 30 sec
72C 1 min
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 MGAT 5 (10 uM)
0.5 ul MGAT 3 (10 uM)
0.5 ul Taq (5 U/ul)
2 ul DNA
--------
25 ul
These primers have been tested in mouse tail DNA preparations to genotype MGAT1 floxed animals.
New feature:
You can add comments on protocols now. Just go to any individual method page and click on
the "Add a comment" link.
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.
...
Read more highlights.
