Mechanically Induced Catalytic Amplification Reaction for Readout of Receptor-Mediated Cellular Forces

• Type of Material: Article
• Authors: Victor Pui-Yan Ma, Emory University; Yang Liu, Emory University; Dr. Kevin Yehl, Emory University; Kornelia Galior, Emory University; Yun Zhang, Emory University; Khalid Salaita, Emory University
• Language: English
• Date: 2016-04-25
• Publisher: Wiley
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
• Final Published Version (URL): http://dx.doi.org/10.1002/anie.201600351
• Title of Journal or Parent Work: Angewandte Chemie International Edition
• ISSN: 1433-7851
• Volume: 55
• Issue: 18
• Start Page: 5488
• End Page: 5492
• Grant/Funding Information: K.S. would like to thank for the financial support from the NIH (R01-GM097399), the Alfred P. Sloan Research Fellowship, the Camille-Dreyfus Teacher-Scholar Award, and the NSF CAREER Award (1350829).
• Keywords: CELLS; mechanically induced catalytic amplification; rolling circle amplification; Science & Technology; MECHANOTRANSDUCTION; drug screening; SURFACE; FOCAL ADHESIONS; PROBES; receptor-mediated forces; DNA-POLYMERASE; Physical Sciences; TENSION; integrins; MOLECULES; INTEGRIN FORCES; Chemistry, Multidisciplinary; Chemistry
• Research Categories: Engineering, Biomedical; Chemistry, General

Abstract

Mechanics play a fundamental role in cell biology, but detecting piconewton (pN) forces is challenging because of a lack of accessible and high throughput assays. A mechanically induced catalytic amplification reaction (MCR) for readout of receptor-mediated forces in cells is described. Mechanically labile DNA duplexes presenting ligands are surface immobilized such that specific receptor forces denature the duplex and thus expose a blocked primer. Amplification of primers is achieved using an isothermal polymerization reaction and quantified by fluorescence readout. As a proof of concept, the assay was used to test the activity of a mechanomodulatory drug and integrin adhesion receptor antibodies. To the best of our knowledge, this is the first example of a catalytic reaction triggered in response to molecular piconewton forces. The MCR may transform the field of mechanobiology by providing a new facile tool to detect receptor specific mechanics with the convenience of the polymerase chain reaction (PCR).