Rapid one-step selection method for generating nucleic acid aptamers: Development of a DNA aptamer against α-bungarotoxin

Lauridsen, Lasse H., Shamaileh, Hadi A., Edwards, Stacey L., Taran, Elena and Veedu, Rakesh N. (2012) Rapid one-step selection method for generating nucleic acid aptamers: Development of a DNA aptamer against α-bungarotoxin. PLoS One, 7 7: e41702.1-e41702.6. doi:10.1371/journal.pone.0041702


Author Lauridsen, Lasse H.
Shamaileh, Hadi A.
Edwards, Stacey L.
Taran, Elena
Veedu, Rakesh N.
Title Rapid one-step selection method for generating nucleic acid aptamers: Development of a DNA aptamer against α-bungarotoxin
Journal name PLoS One   Check publisher's open access policy
ISSN 1932-6203
Publication date 2012-07
Sub-type Article (original research)
DOI 10.1371/journal.pone.0041702
Open Access Status DOI
Volume 7
Issue 7
Start page e41702.1
End page e41702.6
Total pages 6
Place of publication San Francisco, CA, United States
Publisher Public Library of Science
Collection year 2013
Language eng
Formatted 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.
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Article # e41702

 
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Created: Thu, 02 Aug 2012, 16:07:56 EST by Lucy O'Brien on behalf of School of Chemistry & Molecular Biosciences