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Methods Mol Biol. 2014;1103:11-29. doi: 10.1007/978-1-62703-730-3_2.

Screening inhibitory potential of anti-HIV RT RNA aptamers.

Author information

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

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.

PMID:
24318883
DOI:
10.1007/978-1-62703-730-3_2
[Indexed for MEDLINE]

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