转基因
心脏发育
胚胎干细胞
生物
胚胎心脏
电穿孔
转基因
细胞生物学
再生(生物学)
斑马鱼
基因靶向
转基因小鼠
基因
胚胎发生
遗传学
胚胎
生殖技术
作者
Jorge Mañes-García,Leonardo Beccari,Miguel Torres,Oscar H. Ocaña
摘要
The mammalian heart is a complex organ formed during development via highly diverse populations of progenitor cells. The origin, timing of recruitment, and fate of these progenitors are vital for the proper development of this organ. The molecular mechanisms that govern the morphogenesis of the heart are essential for understanding the pathogenesis of congenital heart diseases and embryonic cardiac regeneration. Classical approaches to investigate these mechanisms employed the generation of transgenic mice to assess the function of specific genes during cardiac development. However, mouse transgenesis is a complex, time-consuming process that often cannot be performed to assess the role of specific genes during heart development. To address this, we have developed a protocol for efficient electroporation and culture of mouse embryonic hearts, enabling transient transgenesis to rapidly assess the effect of gain- or loss-of-function of genes involved in cardiac development. Using this methodology, we successfully overexpressed Meis1 in the embryonic heart, with a preference for epicardial cell transfection, demonstrating the capabilities of the technique.
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