PUBLICATION

Bioorthogonal mRNA labeling at the poly(A) tail for imaging localization and dynamics in live zebrafish embryos

Authors
Westerich, K.J., Chandrasekaran, K.S., Gross-Thebing, T., Kueck, N., Raz, E., Rentmeister, A.
ID
ZDB-PUB-210225-2
Date
2020
Source
Chemical science   11: 3089-3095 (Journal)
Registered Authors
Gross-Thebing, Theresa, Raz, Erez, Westerich, Kim
Keywords
none
MeSH Terms
none
PubMed
33623655 Full text @ Chem Sci
Abstract
Live imaging of mRNA in cells and organisms is important for understanding the dynamic aspects underlying its function. Ideally, labeling of mRNA should not alter its structure or function, nor affect the biological system. However, most methods applied in vivo make use of genetically encoded tags and reporters that significantly enhance the size of the mRNA of interest. Alternately, we utilize the 3' poly(A) tail as a non-coding repetitive hallmark to covalently label mRNAs via bioorthogonal chemistry with different fluorophores from a wide range of spectra without significantly changing the size. We demonstrate that the labeled mRNAs can be visualized in cells and zebrafish embryos, and that they are efficiently translated. Importantly, the labeled mRNAs acquired the proper subcellular localization in developing zebrafish embryos and their dynamics could be tracked in vivo.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping