PUBLICATION
Combining near-infrared fluorescence with Brainbow to visualize expression of specific genes within a multicolor context
- Authors
- Cook, Z.T., Brockway, N.L., Tobias, Z.J.C., Pajarla, J., Boardman, I.S., Ippolito, H., Nkombo-Nkoula, S., Weissman, T.A.
- ID
- ZDB-PUB-181227-8
- Date
- 2018
- Source
- Molecular biology of the cell 30(4): 491-505 (Journal)
- Registered Authors
- Weissman, Tamily
- Keywords
- none
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Bone Morphogenetic Protein Receptors/metabolism
- Color
- Embryo, Nonmammalian/metabolism
- Fluorescence
- Gene Expression Regulation*
- Infrared Rays*
- Luminescent Proteins/metabolism*
- Photobleaching
- Zebrafish/embryology
- Zebrafish/genetics*
- Zebrafish/metabolism
- PubMed
- 30586321 Full text @ Mol. Biol. Cell
Citation
Cook, Z.T., Brockway, N.L., Tobias, Z.J.C., Pajarla, J., Boardman, I.S., Ippolito, H., Nkombo-Nkoula, S., Weissman, T.A. (2018) Combining near-infrared fluorescence with Brainbow to visualize expression of specific genes within a multicolor context. Molecular biology of the cell. 30(4):491-505.
Abstract
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 (BMP) receptor within the Brainbow context. The overlay of near-infrared fluorescence tagging 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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping