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Therapeutic Applications of Ribozymes and Riboswitches pp 11–29Cite as

Screening Inhibitory Potential of Anti-HIV RT RNA Aptamers

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1103))

Abstract

Aptamers targeted to HIV reverse transcriptase (RT) have been demonstrated to inhibit RT in biochemical assays and as in cell culture. However, methods employed to date to evaluate viral suppression utilize time-consuming serial passage of infectious HIV in aptamer-expressing stable cell lines. We have established a rapid, transfection-based assay system to effectively examine the inhibitory potential of anti-HIV RT aptamers expressed between two catalytically inactive hammerhead ribozymes. Our system can be altered and optimized for a variety of cloning schemes, and addition of sequences of interest to the cassette is simple and straightforward. When paired with methods to analyze aptamer RNA accumulation and localization in cells and as packaging into pseudotyped virions, the method has a very high level of success in predicting good inhibitors.

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Author information

Authors and Affiliations

  1. Department of Molecular Microbiology & Immunology, University of Missouri, Columbia, MO, USA

    Margaret J. Lange & Donald H. Burke

  2. Department of Biochemistry, University of Missouri, Columbia, MO, USA

    Donald H. Burke

Authors
  1. Margaret J. Lange
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  2. Donald H. Burke
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Editor information

Editors and Affiliations

  1. Department of Biology, Université de Sherbrooke, Québec, Canada

    Daniel Lafontaine

  2. Départment de biologie, Université de Sherbrooke, Sherbrooke, Québec, Canada

    Audrey Dubé

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Lange, M.J., Burke, D.H. (2014). Screening Inhibitory Potential of Anti-HIV RT RNA Aptamers. In: Lafontaine, D., Dubé, A. (eds) Therapeutic Applications of Ribozymes and Riboswitches. Methods in Molecular Biology, vol 1103. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-730-3_2

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  • DOI: https://doi.org/10.1007/978-1-62703-730-3_2

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-729-7

  • Online ISBN: 978-1-62703-730-3

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