Combining near-infrared fluorescence with Brainbow to visualize expression of specific genes within a multicolor context

被引:10
|
作者
Cook, Zoe T. [1 ]
Brockway, Nicole L. [1 ]
Tobias, Zachary J. C. [1 ]
Pajarla, Joy [1 ]
Boardman, Isaac S. [1 ]
Ippolito, Helen [1 ]
Nkoula, Sylvia Nkombo [1 ]
Weissman, Tamily A. [1 ]
机构
[1] Lewis & Clark Coll, Biol Dept, Portland, OR 97219 USA
基金
美国国家科学基金会; 美国国家卫生研究院;
关键词
FAR-RED; BACTERIAL PHYTOCHROMES; HEMATOPOIETIC STEM; HEME OXYGENASE; PROTEINS; CELL; TOOLS; BRIGHT; LIGHT; BACTERIOPHYTOCHROME;
D O I
10.1091/mbc.E18-06-0340
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Fluorescent proteins are a powerful experimental tool, allowing the visualization of gene expression and cellular behaviors in a variety of systems. Multicolor combinations of fluorescent proteins, such as Brainbow, have expanded the range of possible research questions and are useful for distinguishing and tracking cells. The addition of a separately driven color, however, would allow researchers to report expression of a manipulated gene within the multicolor context to investigate mechanistic effects. A far-red or near-infrared protein could be particularly suitable in this context, as these can be distinguished spectrally from Brainbow. We investigated five far-red/near-infrared proteins in zebrafish: TagRFP657, mCardinal, miRFP670, iRFP670, and mIFP. Our results show that both mCardinal and iRFP670 are useful fluorescent proteins for zebrafish expression. We also introduce a new transgenic zebrafish line that expresses Brainbow under the control of the neuroD promoter. We demonstrate that mCardinal can be used to track the expression of a manipulated bone morphogenetic protein receptor within the Brainbow context. The overlay of near-infrared fluorescence onto a Brainbow background defines a clear strategy for future research questions that aim to manipulate or track the effects of specific genes within a population of cells that are delineated using multicolor approaches.
引用
收藏
页码:491 / 505
页数:15
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