Behrouz Asgari Abibeiglou
1 
, Magid Fallahi
1, Mahdi Hesaraki
2, Meghdad Yeganeh
3 , Mohammad Rezaei
1,4, Hossein Baharvand
1,5 , Mohsen Basiri
1,6* , Yaser Tahamtani
1,4* 1 Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran
2 Department of Animal Sciences and Marine Biology, Faculty of Life Science and Biotechnology, Shahid Beheshti University, Tehran Iran
3 School of Biotechnology, College of Science, University of Tehran, Tehran, Iran
4 Department of Basic and Population Based Studies in NCD, Reproductive Epidemiology Research Center, Royan Institute, ACECR, Tehran, Iran
5 Department of Developmental Biology, University of Science and Culture, Tehran, Iran
6 Department of Hematology and Hematopoietic Cell Transplantation (T Cell Therapeutics Research Laboratories), City of Hope Beckman Research Institute and Medical Center, Duarte, CA, USA
Abstract
Introduction: Transgenic ROSA26mT-mG knock-in mice serve as an invaluable tool in developmental and differentiation studies, particularly for tracing and analyzing cell lineages throughout various stages of growth.
Methods: In this study, we employed the CRISPR/Cas9 system to precisely introduce desired genetic material (“ROSA26mT-mG” vector) into the Rosa26 locus of the mouse genome.
Results: Under fluorescent light, ROSA26mT-mG knock-in mice exhibit a red phenotype in the absence of Cre recombinase. However, when crossed with the “UBC-Cre/ER” transgenic mouse, the membrane-targeted tdTomato(mT) gene is excised at the LoxP site, leading to the expression of the membrane-targeted green fluorescent protein (mGFP) gene, thereby changing the mouse's appearance from red to green under fluorescent light.
Conclusion: ROSA26mT-mG knock-in mice can serve as universal lineage tracing models in a diverse array of developmental biology and regenerative medicine studies, enabling researchers to investigate differentiation pathways, and tissue regeneration mechanisms in a clear and visually interpretable manner.