Koehler, Sybille, Braehler, Sebastian, Braun, Fabian ORCID: 0000-0002-4742-050X, Hagmann, Henning, Rinschen, Markus M., Spaeth, Martin R., Hoehne, Martin, Wunderlich, F. Thomas, Schermer, Bernhard ORCID: 0000-0002-5194-9000, Benzing, Thomas and Brinkkoetter, Paul T. (2017). Construction of a viral T2A-peptide based knock-in mouse model for enhanced Cre recombinase activity and fluorescent labeling of podocytes. Kidney Int., 91 (6). S. 1510 - 1518. NEW YORK: ELSEVIER SCIENCE INC. ISSN 1523-1755

Full text not available from this repository.

Abstract

Podocyte injury is a key event in glomerular disease leading to proteinuria and opening the path toward glomerular scarring. As a consequence, glomerular research strives to discover molecular mechanisms and signaling pathways affecting podocyte health. The hNphs2.Cre mouse model has been a valuable tool to manipulate podocyte-specific genes and to label podocytes for lineage tracing and purification. Here we designed a novel podocyte-specific tricistronic Cre mouse model combining codon improved Cre expression and fluorescent cell labeling with mTomato under the control of the endogenous Nphs2 promoter using viral T2A-peptides. Independent expression of endogenous podocin, codon improved Cre, and mTomato was confirmed by immunofluorescence, fluorescent activated cell sorting Nphs2(pod.T2A.ciCre.T2A.mTomato/wild-type) and protein analyses. Nphs2 mice developed normally and did not show any signs of glomerular disease or off-target effects under. basal conditions and in states of disease. Nphs2(pod.T2A.ciCre.T2A.mTomato/wild-type)-mediated gene recombination was superior to conventional hNphs2.Cre mice-mediated gene recombination. Last, we compared Cre efficiency in a disease model by mating Nphs2(pod.T2A.ciCre.T2A.mTomato/wild-type) and hNphs2.Cre mice to Phb2(fl/fl) mice. The podocyte-specific Phb2 knockout by Nphs2(pod.T2A.ciCre.T2A.mTomato/wild-type) mice resulted in an aggravated glomerular injury as compared to a podocyte-specific Phb2 gene deletion triggered by hNphs2.Cre. Thus, we generated the first tricistronic podocyte mouse model combining enhanced Cre recombinase efficiency and fluorescent labeling in podocytes without the need for additional matings with conventional reporter mouse lines.

Item Type: Journal Article
Creators:
CreatorsEmailORCIDORCID Put Code
Koehler, SybilleUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Braehler, SebastianUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Braun, FabianUNSPECIFIEDorcid.org/0000-0002-4742-050XUNSPECIFIED
Hagmann, HenningUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Rinschen, Markus M.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Spaeth, Martin R.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Hoehne, MartinUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Wunderlich, F. ThomasUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Schermer, BernhardUNSPECIFIEDorcid.org/0000-0002-5194-9000UNSPECIFIED
Benzing, ThomasUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Brinkkoetter, Paul T.UNSPECIFIEDUNSPECIFIEDUNSPECIFIED
URN: urn:nbn:de:hbz:38-229285
DOI: 10.1016/j.kint.2016.12.011
Journal or Publication Title: Kidney Int.
Volume: 91
Number: 6
Page Range: S. 1510 - 1518
Date: 2017
Publisher: ELSEVIER SCIENCE INC
Place of Publication: NEW YORK
ISSN: 1523-1755
Language: English
Faculty: Unspecified
Divisions: Unspecified
Subjects: no entry
Uncontrolled Keywords:
KeywordsLanguage
RIBOSOME ENTRY SITES; SIGNAL SEQUENCES; EXPRESSION; CLEAVAGE; VECTORS; BIOLOGY; PROTEIN; COEXPRESSION; POLYPROTEIN; INHIBITIONMultiple languages
Urology & NephrologyMultiple languages
Refereed: Yes
URI: http://kups.ub.uni-koeln.de/id/eprint/22928

Downloads

Downloads per month over past year

Altmetric

Export

Actions (login required)

View Item View Item