Identification of rat Rosa26 locus enables generation of knock-in rat lines ubiquitously expressing tdTomato.

Stem Cells and Development
Toshihiro KobayashiHiromitsu Nakauchi

Abstract

Recent discovery of a method for derivation and culture of germline-competent rat pluripotent stem cells (PSCs) enables generation of transgenic rats or knock-out rats via genetic modification of such PSCs. This opens the way to use rats, as is routine in mice, for analyses of gene functions or physiological features. In mouse or human, one widely used technique to express a gene of interest stably and ubiquitously is to insert that gene into the Rosa26 locus via gene targeting of PSCs. Rosa26 knock-in mice conditionally expressing a reporter or a toxin gene have contributed to tracing or ablation of specific cell lineages. We successfully identified a rat orthologue of the mouse Rosa26 locus. Insertion of tdTomato, a variant of red fluorescent protein, into the Rosa26 locus of PSCs of various rat strains allows ubiquitous expression of tdTomato. Through germline transmission of one Rosa26-tdTomato knock-in embryonic stem cell line, we also obtained tdTomato knock-in rats. These expressed tdTomato ubiquitously throughout their bodies, which indicates that the rat Rosa26 locus conserves functions of its orthologues in mouse and human. The new tools described here (targeting vectors, knock-in PSCs, and rats) should be useful for ...Continue Reading

References

Apr 15, 1997·Proceedings of the National Academy of Sciences of the United States of America·B P ZambrowiczP Soriano
Sep 24, 1999·Developmental Biology·W C KisseberthE P Sandgren
Nov 27, 2007·Nature Biotechnology·Stefan IrionGordon Keller
May 24, 2008·Nature·Qi-Long YingAustin Smith
Nov 10, 2009·Molecular Reproduction and Development·Masumi HirabayashiHiromitsu Nakauchi
Apr 28, 2010·Molecular Reproduction and Development·Masumi HirabayashiHiromitsu Nakauchi
Jul 28, 2010·Proceedings of the National Academy of Sciences of the United States of America·Masaki Kawamata, Takahiro Ochiya
Jul 27, 2011·PloS One·Sanae HamanakaHiromitsu Nakauchi
Apr 14, 2012·Molecular Reproduction and Development·Masumi HirabayashiShinichi Hochi

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Citations

Jan 17, 2016·BMC Biotechnology·Van Trung ChuRalf Kühn
Sep 12, 2015·Molecular Reproduction and Development·Teppei GotoMasumi Hirabayashi
Dec 10, 2013·The Journal of Reproduction and Development·Masumi HirabayashiShinichi Hochi
Apr 30, 2015·Journal of Cardiovascular Translational Research·Michael J FlisterUNKNOWN Rat Genome and Nomenclature Committee
Sep 6, 2014·Stem Cells and Development·Toshihiro KobayashiHiromitsu Nakauchi
Aug 29, 2014·Biology of Reproduction·Tomonari HayamaHiromitsu Nakauchi
Jul 15, 2017·Wiley Interdisciplinary Reviews. Nanomedicine and Nanobiotechnology·Anna V Naumova, Greetje Vande Velde
Mar 2, 2018·Tissue Engineering. Part C, Methods·Brian C SyverudLisa M Larkin
Mar 23, 2018·G3 : Genes - Genomes - Genetics·Mengjing LiDaxin Pang
Feb 1, 2020·Development·Toshihiro KobayashiMasumi Hirabayashi
Aug 22, 2017·The Journal of Reproduction and Development·Masumi HirabayashiHiromitsu Nakauchi
Feb 7, 2019·Nature Communications·Teppei GotoMasumi Hirabayashi
Mar 21, 2019·Animal Models and Experimental Medicine·Shun LiXiao-Hui Zhou
Jul 29, 2017·Mammalian Genome : Official Journal of the International Mammalian Genome Society·Stephen MeekTom Burdon
Oct 18, 2018·Scientific Reports·Tomoyuki YamaguchiHiromitsu Nakauchi
Apr 12, 2018·American Journal of Physiology. Renal Physiology·Xiaohui GongXingqun Liang
Dec 14, 2019·Journal of Biological Engineering·Long XieYangqing Lu
Jul 12, 2018·Scientific Reports·Ming WangYunping Dai
May 8, 2021·Frontiers in Genetics·Vanessa ChenouardIgnacio Anegon
Jul 22, 2021·Biochemistry. Biokhimii︠a︡·Vladimir V Sherstyuk, Suren M Zakian
Oct 21, 2021·Biology of Reproduction·Nima DehdilaniHesam Dehghani

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Methods Mentioned

BETA
transgenic
gene-trap
PCR
flow cytometry
FACS
fluorescence activated cell sorting

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