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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2019
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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. |
format | Online Article Text |
id | pubmed-6594444 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
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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