Fraser Stuart T, Isern Joan, Baron Margaret H
Division of Hematology and Medical Oncology, Department of Medicine, Mount Sinai School of Medicine, New York, USA.
Methods Enzymol. 2010;476:403-27. doi: 10.1016/S0076-6879(10)76022-5.
The use of fluorescent reporter proteins such as GFP, RFP, and their variants to tag and track cells within the embryo has revolutionized developmental biology. Expression of these proteins within restricted populations has been achieved through the use of lineage-specific regulatory elements. This approach has proven especially powerful in the hematopoietic system, where it has been possible to monitor the generation, expansion, maturation, and migration of primitive erythroid cells, macrophages, and megakaryocytes during embryogenesis at unprecedented resolution. Such analyses have provided novel insights into the development of these lineages. In this chapter, we discuss the design considerations and methodologies involved in the production and analysis of transgenic mouse lines in which fluorescent reporters are expressed in the hematopoietic system of the mouse embryo.
利用绿色荧光蛋白(GFP)、红色荧光蛋白(RFP)及其变体等荧光报告蛋白对胚胎内的细胞进行标记和追踪,彻底改变了发育生物学。通过使用谱系特异性调控元件,已实现这些蛋白在特定细胞群体中的表达。这种方法在造血系统中已证明特别有效,利用它能够以前所未有的分辨率监测胚胎发育过程中原始红细胞、巨噬细胞和巨核细胞的产生、扩增、成熟和迁移。此类分析为这些细胞谱系的发育提供了新的见解。在本章中,我们将讨论在小鼠胚胎造血系统中表达荧光报告基因的转基因小鼠品系的构建及分析所涉及的设计考量和方法。