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

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 distinguis...

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Autores principales: Cook, Zoe T., Brockway, Nicole L., Tobias, Zachary J. C., Pajarla, Joy, Boardman, Isaac S., Ippolito, Helen, Nkombo Nkoula, Sylvia, Weissman, Tamily A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The American Society for Cell Biology 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6594444/
https://www.ncbi.nlm.nih.gov/pubmed/30586321
http://dx.doi.org/10.1091/mbc.E18-06-0340
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author Cook, Zoe T.
Brockway, Nicole L.
Tobias, Zachary J. C.
Pajarla, Joy
Boardman, Isaac S.
Ippolito, Helen
Nkombo Nkoula, Sylvia
Weissman, Tamily A.
author_facet Cook, Zoe T.
Brockway, Nicole L.
Tobias, Zachary J. C.
Pajarla, Joy
Boardman, Isaac S.
Ippolito, Helen
Nkombo Nkoula, Sylvia
Weissman, Tamily A.
author_sort Cook, Zoe T.
collection PubMed
description 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.
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spelling pubmed-65944442019-07-05 Combining near-infrared fluorescence with Brainbow to visualize expression of specific genes within a multicolor context Cook, Zoe T. Brockway, Nicole L. Tobias, Zachary J. C. Pajarla, Joy Boardman, Isaac S. Ippolito, Helen Nkombo Nkoula, Sylvia Weissman, Tamily A. Mol Biol Cell Articles 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. The American Society for Cell Biology 2019-02-15 /pmc/articles/PMC6594444/ /pubmed/30586321 http://dx.doi.org/10.1091/mbc.E18-06-0340 Text en © 2019 Cook et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. http://creativecommons.org/licenses/by-nc-sa/3.0 This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License.
spellingShingle Articles
Cook, Zoe T.
Brockway, Nicole L.
Tobias, Zachary J. C.
Pajarla, Joy
Boardman, Isaac S.
Ippolito, Helen
Nkombo Nkoula, Sylvia
Weissman, Tamily A.
Combining near-infrared fluorescence with Brainbow to visualize expression of specific genes within a multicolor context
title Combining near-infrared fluorescence with Brainbow to visualize expression of specific genes within a multicolor context
title_full Combining near-infrared fluorescence with Brainbow to visualize expression of specific genes within a multicolor context
title_fullStr Combining near-infrared fluorescence with Brainbow to visualize expression of specific genes within a multicolor context
title_full_unstemmed Combining near-infrared fluorescence with Brainbow to visualize expression of specific genes within a multicolor context
title_short Combining near-infrared fluorescence with Brainbow to visualize expression of specific genes within a multicolor context
title_sort combining near-infrared fluorescence with brainbow to visualize expression of specific genes within a multicolor context
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6594444/
https://www.ncbi.nlm.nih.gov/pubmed/30586321
http://dx.doi.org/10.1091/mbc.E18-06-0340
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