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生成用于肾脏发育研究的基因镶嵌小鼠胚胎或类器官。

Generating Genetic Mosaic Mouse Embryos or Organoids for Studies of Kidney Development.

作者信息

Costantini Frank

机构信息

Department of Genetics and Development, 1418 Hammer Health Sciences Center, Columbia University, New York, NY, USA.

出版信息

Methods Mol Biol. 2019;1926:3-21. doi: 10.1007/978-1-4939-9021-4_1.

Abstract

For studies of gene function during development, it can be very useful to generate mosaic embryos in which a small subset of cells in a given cell lineage lacks a gene of interest and carries a marker that allows the mutant cells to be specifically visualized and compared to wild-type cells. Several methods have been used to generate genetically mosaic mouse kidneys for such studies. These include (1) chimeric embryos generated using embryonic stem cells, (2) chimeric renal organoids generated by dissociation and reaggregation of the fetal kidneys, (3) generation of a knockout allele with a built-in reporter gene, (4) mosaic analysis with double markers (MADM), and (5) mosaic mutant analysis with spatial and temporal control of recombination (MASTR). In this chapter, these five methods are described, and their advantages and disadvantages are discussed.

摘要

对于发育过程中基因功能的研究,生成嵌合胚胎可能非常有用,在这种胚胎中,特定细胞谱系中的一小部分细胞缺乏感兴趣的基因,并带有一种标记物,使突变细胞能够被特异性可视化,并与野生型细胞进行比较。已经使用了几种方法来生成用于此类研究的基因嵌合小鼠肾脏。这些方法包括:(1) 使用胚胎干细胞生成的嵌合胚胎;(2) 通过胎儿肾脏解离和重新聚集生成的嵌合肾类器官;(3) 带有内置报告基因的敲除等位基因的生成;(4) 双标记嵌合分析 (MADM);以及(5) 具有重组时空控制的嵌合突变分析 (MASTR)。在本章中,将描述这五种方法,并讨论它们的优缺点。

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