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Genesis. 2017 Aug;55(8). doi: 10.1002/dvg.23041. Epub 2017 Jun 15.

Generation of Nkx2-5/CreER transgenic mice for inducible Cre expression in developing hearts.

Ranjbarvaziri S1,2,3, Park S1,2,3, Nguyen NB1,2,3, Gilmore WB1,2, Zhao P1,2,3, Ardehali R1,2,3.

Author information

1
Division of Cardiology, Department of Internal Medicine, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, California, 90095.
2
Eli and Edythe Broad Center for Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, California, 90095.
3
Molecular, Cellular and Integrative Physiology Graduate Program, University of California, Los Angeles, Los Angeles, California, 90095.

Abstract

Nkx2-5 is a homeobox-containing transcriptional regulator that serves as one of the earliest markers of cardiac lineage commitment. To study the role of Nkx2-5-expressing progenitors at specific time points in cardiac development, we have generated a novel and inducible NKX2-5 mouse line by knocking in a CreER cassette into the Nkx2-5 genomic locus, while preserving the endogenous Nkx2-5 gene to avoid haploinsufficiency. We evaluated the specificity and efficiency of CreER activity after 4-OHT injection by crossing Nkx2-5CreER/+ mice with a Rosa26tdT/+ reporter strain. Our immunohistochemistry results confirmed Cre-induced tdTomato expression specifically in cells expressing Nkx2-5. These cells were mainly cardiomyocytes and were observed in the embryonic heart as early as day 9.5. Additionally, quantitative polymerase chain reaction on postnatal hearts showed enriched expression of Nkx2-5 in isolated tdTomato-expressing cells. No tdTomato expression was observed in Nkx2-5CreER/+ ;Rosa26tdT/+ mice in the absence of 4-OHT, confirming the inducible nature of CreER activity. The Nkx2-5/CreER mouse model described in this article will serve as an invaluable tool to trace myocardial lineage and to temporally induce genetic manipulation in a selective population of cardiac progenitors during embryonic development and in the adult heart.

KEYWORDS:

Nkx2-5; cardiac development; inducible CreER recombinase

PMID:
28589709
PMCID:
PMC5568924
DOI:
10.1002/dvg.23041
[Indexed for MEDLINE]
Free PMC Article

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