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Construction of a viral T2A-peptide based knock-in mouse model for enhanced Cre recombinase activity and fluorescent labeling of podocytes

Standard

Construction of a viral T2A-peptide based knock-in mouse model for enhanced Cre recombinase activity and fluorescent labeling of podocytes. / Koehler, Sybille; Brähler, Sebastian; Braun, Fabian; Hagmann, Henning; Rinschen, Markus M; Späth, Martin R; Höhne, Martin; Wunderlich, F Thomas; Schermer, Bernhard; Benzing, Thomas; Brinkkoetter, Paul T.

In: KIDNEY INT, Vol. 91, No. 6, 06.2017, p. 1510-1517.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Koehler, S, Brähler, S, Braun, F, Hagmann, H, Rinschen, MM, Späth, MR, Höhne, M, Wunderlich, FT, Schermer, B, Benzing, T & Brinkkoetter, PT 2017, 'Construction of a viral T2A-peptide based knock-in mouse model for enhanced Cre recombinase activity and fluorescent labeling of podocytes', KIDNEY INT, vol. 91, no. 6, pp. 1510-1517. https://doi.org/10.1016/j.kint.2016.12.011

APA

Koehler, S., Brähler, S., Braun, F., Hagmann, H., Rinschen, M. M., Späth, M. R., Höhne, M., Wunderlich, F. T., Schermer, B., Benzing, T., & Brinkkoetter, P. 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), 1510-1517. https://doi.org/10.1016/j.kint.2016.12.011

Vancouver

Koehler S, Brähler S, Braun F, Hagmann H, Rinschen MM, Späth MR et al. Construction of a viral T2A-peptide based knock-in mouse model for enhanced Cre recombinase activity and fluorescent labeling of podocytes. KIDNEY INT. 2017 Jun;91(6):1510-1517. https://doi.org/10.1016/j.kint.2016.12.011

Bibtex

@article{b796530dd0a24ebf833e600cd244f0fd,
title = "Construction of a viral T2A-peptide based knock-in mouse model for enhanced Cre recombinase activity and fluorescent labeling of podocytes",
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 and protein analyses. Nphs2pod.T2A.ciCre.T2A.mTomato/wild-typemice developed normally and did not show any signs of glomerular disease or off-target effects under basal conditions and in states of disease. Nphs2pod.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 Nphs2pod.T2A.ciCre.T2A.mTomato/wild-typeand hNphs2.Cre mice to Phb2fl/flmice. The podocyte-specific Phb2 knockout by Nphs2pod.T2A.ciCre.T2A.mTomato/wild-typemice 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.",
keywords = "Journal Article",
author = "Sybille Koehler and Sebastian Br{\"a}hler and Fabian Braun and Henning Hagmann and Rinschen, {Markus M} and Sp{\"a}th, {Martin R} and Martin H{\"o}hne and Wunderlich, {F Thomas} and Bernhard Schermer and Thomas Benzing and Brinkkoetter, {Paul T}",
note = "Copyright {\textcopyright} 2016 International Society of Nephrology. Published by Elsevier Inc. All rights reserved.",
year = "2017",
month = jun,
doi = "10.1016/j.kint.2016.12.011",
language = "English",
volume = "91",
pages = "1510--1517",
journal = "KIDNEY INT",
issn = "0085-2538",
publisher = "NATURE PUBLISHING GROUP",
number = "6",

}

RIS

TY - JOUR

T1 - Construction of a viral T2A-peptide based knock-in mouse model for enhanced Cre recombinase activity and fluorescent labeling of podocytes

AU - Koehler, Sybille

AU - Brähler, Sebastian

AU - Braun, Fabian

AU - Hagmann, Henning

AU - Rinschen, Markus M

AU - Späth, Martin R

AU - Höhne, Martin

AU - Wunderlich, F Thomas

AU - Schermer, Bernhard

AU - Benzing, Thomas

AU - Brinkkoetter, Paul T

N1 - Copyright © 2016 International Society of Nephrology. Published by Elsevier Inc. All rights reserved.

PY - 2017/6

Y1 - 2017/6

N2 - 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 and protein analyses. Nphs2pod.T2A.ciCre.T2A.mTomato/wild-typemice developed normally and did not show any signs of glomerular disease or off-target effects under basal conditions and in states of disease. Nphs2pod.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 Nphs2pod.T2A.ciCre.T2A.mTomato/wild-typeand hNphs2.Cre mice to Phb2fl/flmice. The podocyte-specific Phb2 knockout by Nphs2pod.T2A.ciCre.T2A.mTomato/wild-typemice 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.

AB - 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 and protein analyses. Nphs2pod.T2A.ciCre.T2A.mTomato/wild-typemice developed normally and did not show any signs of glomerular disease or off-target effects under basal conditions and in states of disease. Nphs2pod.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 Nphs2pod.T2A.ciCre.T2A.mTomato/wild-typeand hNphs2.Cre mice to Phb2fl/flmice. The podocyte-specific Phb2 knockout by Nphs2pod.T2A.ciCre.T2A.mTomato/wild-typemice 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.

KW - Journal Article

U2 - 10.1016/j.kint.2016.12.011

DO - 10.1016/j.kint.2016.12.011

M3 - SCORING: Journal article

C2 - 28187984

VL - 91

SP - 1510

EP - 1517

JO - KIDNEY INT

JF - KIDNEY INT

SN - 0085-2538

IS - 6

ER -