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MNAzymes, a Versatile New Class of Nucleic Acid Enzymes That Can Function as Biosensors and Molecular Switches

Elisa Mokany*^†, Simon M. Bone^†, Paul E. Young^‡, Tram B. Doan^‡ and Alison V. Todd^†

SpeeDx Pty Ltd, Eveleigh, New South Wales, Australia 2015, and Victor Chang Cardiac Research Institute, Darlinghurst, New South Wales, Australia 2010

J. Am. Chem. Soc., 2010, 132 (3), pp 1051–1059

DOI: 10.1021/ja9076777

Publication Date (Web): December 28, 2009

elisam@speedx.com.au, †

SpeeDx Pty Ltd.

, ‡

Victor Chang Cardiac Research Institute.

Section:

Enzymes

Abstract

To increase the versatility and utility of nucleic acid enzymes, we developed multicomponent complexes, known as MNAzymes, which produce amplified “output” signals in response to specific “input” signals. Multiple oligonucleotide partzymes assemble into active MNAzymes only in the presence of an input assembly facilitator such as a target nucleic acid. Once formed, MNAzymes catalytically modify a generic substrate, generating an amplified output signal that heralds the presence of the target while leaving the target intact. We demonstrated several applications including sensitive, isothermal target detection; discrimination of polymorphisms; and highly specific monitoring of real-time polymerase chain reaction (PCR). Furthermore, we showed their capacity to function as molecular switches and to work in series to create a molecular cascade. The modular nature of MNAzymes, together with the separation of input and output functionalities, provides potential for their integration into diverse devices such as diagnostic biosensors, molecular computers, and/or nanoscale machines.

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