Rapid One-Step Selection Method for Generating Nucleic Acid Aptamers: Development of a DNA Aptamer against α-Bungarotoxin

L.H. Lauridsen; H.A. Shamaileh; S.L. Edwards; E. Taran; R.N. Veedu

doi:10.1371/journal.pone.0041702

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Rapid One-Step Selection Method for Generating Nucleic Acid Aptamers: Development of a DNA Aptamer against α-Bungarotoxin

Journal article

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Peer reviewed

Rapid One-Step Selection Method for Generating Nucleic Acid Aptamers: Development of a DNA Aptamer against α-Bungarotoxin

L.H. Lauridsen, H.A. Shamaileh, S.L. Edwards, E. Taran and R.N. Veedu

PloS one, Vol.7(7), e41702

2012

DOI: https://doi.org/10.1371/journal.pone.0041702

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Abstract

Background Nucleic acids based therapeutic approaches have gained significant interest in recent years towards the development of therapeutics against many diseases. Recently, research on aptamers led to the marketing of Macugen®, an inhibitor of vascular endothelial growth factor (VEGF) for the treatment of age related macular degeneration (AMD). Aptamer technology may prove useful as a therapeutic alternative against an array of human maladies. Considering the increased interest in aptamer technology globally that rival antibody mediated therapeutic approaches, a simplified selection, possibly in one-step, technique is required for developing aptamers in limited time period. Principal Findings Herein, we present a simple one-step selection of DNA aptamers against α-bungarotoxin. A toxin immobilized glass coverslip was subjected to nucleic acid pool binding and extensive washing followed by PCR enrichment of the selected aptamers. One round of selection successfully identified a DNA aptamer sequence with a binding affinity of 7.58 µM. Conclusion We have demonstrated a one-step method for rapid production of nucleic acid aptamers. Although the reported binding affinity is in the low micromolar range, we believe that this could be further improved by using larger targets, increasing the stringency of selection and also by combining a capillary electrophoresis separation prior to the one-step selection. Furthermore, the method presented here is a user-friendly, cheap and an easy way of deriving an aptamer unlike the time consuming conventional SELEX-based approach. The most important application of this method is that chemically-modified nucleic acid libraries can also be used for aptamer selection as it requires only one enzymatic step. This method could equally be suitable for developing RNA aptamers.

Details

Title: Rapid One-Step Selection Method for Generating Nucleic Acid Aptamers: Development of a DNA Aptamer against α-Bungarotoxin
Authors/Creators: L.H. Lauridsen (Author/Creator) - The University of Queensland
H.A. Shamaileh (Author/Creator) - The University of Queensland
S.L. Edwards (Author/Creator) - The University of Queensland
E. Taran (Author/Creator) - Australian National Fabrication Facility
R.N. Veedu (Author/Creator) - The University of Queensland
Publication Details: PloS one, Vol.7(7), e41702
Publisher: Public Library of Science
Identifiers: 991005539979807891
Copyright: © 2012 Lauridsen et al.
Murdoch Affiliation: Murdoch University
Language: English
Resource Type: Journal article

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Collaboration types: Domestic collaboration; International collaboration
Citation topics: 2 Chemistry; 2.145 Biosensors; 2.145.243 Nanobiosensors
Web Of Science research areas: Biochemistry & Molecular Biology
ESI research areas: Biology & Biochemistry