基因示踪


基因示踪是基因修饰小鼠的一种重要应用,通过基因敲入的形式,将外源的示踪基因敲入到内源基因中,从而可以对基因进行标记和示踪。

可以达到以下3类示踪目的:

蛋白的亚定位

可将reporter基因敲入到小鼠内源基因的3'端使与内源基因融合表达,通过观察reporter基因的定位确定内源蛋白的细胞亚定位,如下图所示:

微信图片_20200908153152.jpg

蛋白的表达谱

可将reporter基因敲入到小鼠内源基因的3'端,对內源基因进行标记或者替代表达,通过观察reporter基因的表达了解内源性蛋白的表达情况,例如:是否表达、表达量、表达组织分布),如下图所示:

微信图片_20200908153207.png

细胞的发育谱系

利用Rosa26-LSL-reporter示踪小鼠,与特定细胞Marker基因敲入CreER的工具小鼠交配,在子代小鼠中使特定类型细胞被永久标记上reporter荧光,便能通过观察这些被标记上的荧光信号来追踪某类特定细胞及其后代所有细胞的增殖、分化以及迁移等活动,为研究细胞命运及谱系提供有力手段。如下图所示:

image.png


遗传谱系示踪应用案例

Enhancing the precision of genetic lineage tracing using dual recombinases


更多谱系示踪相关文献

发表年份

文章题目

发表期刊

2018/1 Genetic Fate Mapping Defines the Vascular Potential of Endocardial Cells in the Adult Heart Circ Res

2017/11

Enhancing the precision of genetic lineage tracing using dual recombinases

Nat Med

2017/8

Fibroblasts in an endocardial fibroelastosis disease model mainly originate from mesenchymal derivatives of 

epicardium

Cell Res

2017/7

Identification of a hybrid myocardial zone in the mammalian heart after birth.

Nat Commun

2017/6

Preexisting endothelial cells mediate cardiac neovascularization after injury

J Clin Invest

2016/11

Mfsd2a+ hepatocytes repopulate the liver during injury and regeneration

Nat Commun

2016/6

Endocardium Minimally Contributes to Coronary  Endothelium in the Embryonic Ventricular Free Walls

Circ Res

2016/3

Genetic lineage tracing identifies endocardial origin of liver vasculature

Nat Genet

2016/1

Endocardium Contributes to Cardiac Fat

Circ Res

2016/1

Genetic lineage tracing identifies in situ Kit-expressing cardiomyocytes

Cell Res

2015/7

c-kit(+) cells adopt vascular endothelial but not epithelial cell fates during lung maintenance and repair

Nat Med


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