Journal
NEPHROLOGY DIALYSIS TRANSPLANTATION
Volume 25, Issue 7, Pages 2120-2124Publisher
OXFORD UNIV PRESS
DOI: 10.1093/ndt/gfq029
Keywords
CreER(T2); Cre recombinase; inducible conditional knockout mice; tamoxifen
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Funding
- Japanese Ministry of Education, Culture, Sports, Science and Technology
- Japanese Ministry of Health, Labour and Welfare, Smoking Research Foundation
- Salt Science Research Foundation
- Grants-in-Aid for Scientific Research [22590910] Funding Source: KAKEN
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Methods. To introduce a temporal control in the genetic experiments targeting the podocyte, we constructed tamoxifen-inducible Cre recombinase (CreER(T2)) transgenic mice under the control of podocyte-specific promoter, 2.5-kb fragment of the human podocin (NPHS2) gene. The specificity and efficiency of Cre activity were examined by crossing NPHS2-CreER(T2) with the ROSA26 reporter (R26R) mouse in which a floxed-stop cassette has been placed upstream of the beta-galactosidase gene. Four-week-old double-mutant mice (NPHS2-CreER(T2)/R26R) were intraperitoneally administered with 0.5 mg of 4-hydroxytamoxifen (4-OHT) for three consecutive days. Results. NPHS2-CreER(T2)/R26R treated with 4-OHT expressed beta-galactosidase specifically in 85% of the podocytes in glomeruli. Expression of Cre recombinase mRNA was mostly restricted to the kidney, especially in glomeruli. Conclusions. In conclusion, we have successfully generated podocyte-specific inducible Cre transgenic mice by tamoxifen administration. These mice allow us to disrupt the genes specifically in the podocytes after birth.
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