Ready-to-use Aptamer Biosensors for DNT and RDX

Abstract

Aptamers are nucleic acid-based reagents that bind to target molecules with high affinity and specificity. However, methods for generating aptamers from random combinatorial libraries (e.g., SELEX) are often labor-intensive and time-consuming. To address this problem, we have recently demonstrated the microfluidic SELEX (M-SELEX) technology that can accelerate aptamer isolation by enabling highly stringent selection conditions through the use of very small amounts of target molecules (X. Lou et a/, PNAS 2009). In this work, we utilize the M-SELEX technology to generate a novel class of Self-Reporting Aptamers (SRAs) which are capable of a) specifically bind to their target, b) undergo conformational change triggered by this binding event, and c) activate a DNA enzyme moiety within the aptamer structure to fluorescently report the binding event. This technique is especially useful for the detection of small molecules because it does not require immobilization of the target molecules on solid support. As a proof of concept, in the last reporting period, we have demonstrated the isolation of SRAs that specifically bind to a target protein with low nanomolar affinity (S. Oh eta/., in review), and we report our progress in generating SRAs against small molecule targets.

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Document Details

Document Type
Technical Report
Publication Date
Sep 29, 2011
Accession Number
ADA551279

Entities

People

  • Hyonsok T. Soh
  • Yao Xiao

Organizations

  • University of California, Santa Barbara

Tags

Communities of Interest

  • Materials and Manufacturing Processes

DTIC Thesaurus Topics

  • Analytical Chemistry
  • Antisense Elements (Genetics)
  • Biosensors
  • Chemical Reactions
  • Chemistry
  • Detection
  • Detectors
  • Dna Sequence Analysis
  • Fluorescence
  • Genetic Structures
  • Magnetic Separation
  • Molecules
  • Nucleic Acids
  • Polymerase Chain Reaction
  • Proteins
  • Sequence Analysis
  • Small Molecules

Fields of Study

  • Chemistry

Readers

  • Molecular Genetics

Technology Areas

  • Biotechnology