Abstract
In situ hybridization allows the detection of nucleic acid sequences in fixed cells and tissues. The gelatinous nature of cnidarians and Hydractinia demands extensive and exhausting protocols to detect RNA transcripts with traditional methods (e.g., colorimetric in situ hybridization). Signal amplification by exchange reaction (SABER) fluorescence in situ hybridization (FISH) enables simplifying and multiplex imaging of RNA targets in a rapid and cost-effective manner. In one enzymatic reaction, SABER-FISH uses a strand-displacing polymerase and catalytic DNA hairpin to generate FISH probes with adjustable signal amplification, allowing highly sensitive detection of nucleic acids and reducing the number of required probes. Here I describe the methodology to detect transcripts within the cells of Hydractinia by SABER-FISH in whole-mount samples.
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Acknowledgments
I thank members of the Frank lab for discussions and advice. Confocal images were taken at the Centre for Microscopy and Imaging Core Facility at University of Galway. MSS is a Human Frontier Science Program Long-Term Postdoctoral Fellow (grant no. LT000756/2020-L).
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Salinas-Saavedra, M. (2024). SABER-FISH in Hydractinia. In: Haimovich, G. (eds) Fluorescence In Situ Hybridization (FISH). Methods in Molecular Biology, vol 2784. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3766-1_5
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DOI: https://doi.org/10.1007/978-1-0716-3766-1_5
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