2nd Epigenetics and Bioengineering Conference (EpiBio 2018)
Exponential Fluorescent Amplification of Individual Nucleic Acids Using Clampfish Probes
Authors
Sara H. Rouhanifard - Presenter, University of Pennsylvania
Ian A. Mellis, University of Pennsylvania
Margaret Dunagin, University of Pennsylvania
Sareh Bayatpour, University of Pennsylvania
Connie L. Jiang, University of Pennsylvania
Ian Dardani, University of Pennsylvania
Orsolya Symmons, University of Pennsylvania
Benjamin Emert, University of Pennsylvania
Eduardo Torre, University of Pennsylvania
Allison Cote, University of Pennsylvania
Alessandra Sullivan, Altius Institute
John Stamatoyannopoulos, Altius Institute
Arjun Raj, University of Pennsylvania
Non-enzymatic, high-gain signal amplification methods with single-cell, single-molecule resolution are in great need. We present click-amplifying FISH (clampFISH) for the fluorescent detection of nucleic acids that combines the specificity of oligonucleotides with bioorthogonal click chemistry in order to achieve high specificity and extremely high-gain (>400x) signal amplification in single cells. We show that clampFISH signal enables detection of RNA species with low magnification microscopy and separation of cells by RNA levels via flow cytometry. Additionally, we show that the modular design of clampFISH probes enables multiplexing of RNA and DNA, that the locking mechanism prevents probe detachment in expansion microscopy, and that clampFISH can be applied to tissue samples.