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Publication : Cre recombinase microinjection for single-cell tracing and localised gene targeting.

First Author  Sendra M Year  2023
Journal  Development Volume  150
Issue  3 PubMed ID  36734327
Mgi Jnum  J:338272 Mgi Id  MGI:7438087
Doi  10.1242/dev.201206 Citation  Sendra M, et al. (2023) Cre recombinase microinjection for single-cell tracing and localised gene targeting. Development 150(3):dev201206
abstractText  Tracing and manipulating cells in embryos are essential to understand development. Lipophilic dye microinjections, viral transfection and iontophoresis have been key to map the origin of the progenitor cells that form the different organs in the post-implantation mouse embryo. These techniques require advanced manipulation skills and only iontophoresis, a demanding approach of limited efficiency, has been used for single-cell labelling. Here, we perform lineage tracing and local gene ablation using cell-permeant Cre recombinase (TAT-Cre) microinjection. First, we map the fate of undifferentiated progenitors to the different heart chambers. Then, we achieve single-cell recombination by titrating the dose of TAT-Cre, which allows clonal analysis of nascent mesoderm progenitors. Finally, injecting TAT-Cre to Mycnflox/flox embryos in the primitive heart tube revealed that Mycn plays a cell-autonomous role in maintaining cardiomyocyte proliferation. This tool will help researchers identify the cell progenitors and gene networks involved in organ development, helping to understand the origin of congenital defects.
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