Rosa26-rtTA-IRES-EGFP transgenic mouse

This mouse model was generated by Nagy and colleagues. It can be used to inducibly expression your favorite transgene by Tet-on system or as a reporter mouse strain to characterize promoter specificity. This mouse can be obtained through Jackson labs.

Ref: Belteki G. et al. Nucleic Acids Research 2005.

ROSA 5:

GAG TTC TCT GCT GCC TCC TG

ROSA 3:

CGA GGC GGA TAC AAG CAA TA

RTTA 3:

AAG ACC GCG AAG AGT TTG TC

ROSA 5 and ROSA 3 amplify a fragment of 322 bp from WT allele. ROSA 5 and RTTA 3 amplify a fragment of 215 bp from knockin allele.

Recommended condition:

94C 5 min
94C 30 sec
60C 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 ROSA 5 (10 uM)
0.5 ul ROSA 3 or RTTA 3 (10 uM)
0.5 ul Taq (5 U/ul)
2 ul DNA
--------
25 ul

These primers have been tested tail DNA preparations.


Comments

Add a comment
Site News
New feature:
You can add comments on protocols now. Just go to any individual method page and click on the "Add a comment" link.

Highlight of the month
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.