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Construction and Analysis of CreERT2 Knock-in Mouse for Genetic Labling of Isl1+ Cardiac Progenitor Cells by CRISPR/Cas9 Technology
Zhou Yunhe1,2, Gu Xiaowen1, Yang Hua1, Huang Dandan1, Fei Jian1,3*
1School of Life Sciences and Technology, Tongji University, Shanghai 200092, China; 2Department of Physical Education,
Tongji University, Shanghai 200092, China; 3Shanghai Research Center for Model Organisms, Shanghai 200092, China
Abstract: Cardiomyocytes in adult mammals retain a limited ability to proliferate in response to specific stimuli， but little is known about the origin of proliferating cells. LIM-homeodomain transcription factor islet- 1(Isl1)-expressing cells are used to study the optimal endogenous progenitor cells in cardiac regeneration. Using the CRISPR/Cas9 genomic editing technology， we constructed Isl1-CreERT2 knock-in mouse model which harboringan CreERT2 cassette down steam of the Isl1 promoter. We crossed Isl1-CreERT2 males with Rosa26-loxP-neo-loxPLacZ transgenic reporter mice (Rosa26-lacZ+) and analyzed Isl1+ positive cells by X-Gal staining. The results showed that Tamoxifen-Inducible Cre/loxP recombination， as depicted by blue cells， existed in heart sinoatrial node， cardiac ganglia， the aortic arch and pulmonary roots in adult mice. Isl1 expression profile was corresponding with previous research. By this work， we established Isl1-CreERT2 knock-in mouse model successfully， and the model provides a useful tool for tracing cardiac progenitor cells in cardiac regeneration. This study will be helpful for understanding and treating cardiovascular diseases， clinical medicine and sports rehabilitation medicine.
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