Comparative assessment of fluorescent transgene methods for quantitative imaging in human cells

被引：37

作者：

Mahen, Robert ^{[1
]}

Koch, Birgit ^{[1
]}

Wachsmuth, Malte ^{[1
]}

Politi, Antonio Z. ^{[1
]}

Perez-Gonzalez, Alexis ^{[1
]}

Mergenthaler, Julia ^{[1
]}

Cai, Yin ^{[1
]}

Ellenberg, Jan ^{[1
]}

机构：

[1] European Mol Biol Lab, D-69117 Heidelberg, Germany

来源：

MOLECULAR BIOLOGY OF THE CELL | 2014年 / 25卷 / 22期

基金：

英国惠康基金;

关键词：

LIVING CELLS; COMPLEX; CYTOKINESIS; GENOME; YEAST; GENE;

D O I：

10.1091/mbc.E14-06-1091

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

Fluorescence tagging of proteins is a widely used tool to study protein function and dynamics in live cells. However, the extent to which different mammalian transgene methods faithfully report on the properties of endogenous proteins has not been studied comparatively. Here we use quantitative live-cell imaging and single-molecule spectroscopy to analyze how different transgene systems affect imaging of the functional properties of the mitotic kinase Aurora B. We show that the transgene method fundamentally influences level and variability of expression and can severely compromise the ability to report on endogenous binding and localization parameters, providing a guide for quantitative imaging studies in mammalian cells.

引用

收藏

页码：3610 / 3618

页数：9

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