Poly-Target Selection Identifies Broad-Spectrum RNA Aptamers
| Autor: | Jonathan L. Chang, Donald H. Burke, Phuong D.M. Nguyen, Margaret J. Lange, Khalid K. Alam, Andrew W. Sawyer |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Mutation rate cross-reactivity Aptamer Population Mutant selection Computational biology Biology 010402 general chemistry 01 natural sciences Article law.invention Broad spectrum 03 medical and health sciences RNA Aptamers law Drug Discovery reverse transcriptase education 030304 developmental biology 0303 health sciences education.field_of_study drug resistance 030102 biochemistry & molecular biology SELEX lcsh:RM1-950 RNA aptamer HIV bioinformatics broad-spectrum inhibitors Phenotype Reverse transcriptase 3. Good health 0104 chemical sciences 030104 developmental biology lcsh:Therapeutics. Pharmacology Viral replication Recombinant DNA Molecular Medicine Systematic evolution of ligands by exponential enrichment |
| Zdroj: | Molecular Therapy: Nucleic Acids, Vol 13, Iss, Pp 605-619 (2018) Molecular Therapy. Nucleic Acids |
| ISSN: | 2162-2531 |
| Popis: | Aptamer selections often yield distinct subpopulations, each with unique phenotypes that can be leveraged for specialized applications. Although most selections aim to attain ever higher specificity, we sought to identify aptamers that recognize increasingly divergent primate lentiviral reverse transcriptases (RTs). We hypothesized that aptamer subpopulations in libraries pre-enriched against a single RT may exhibit broad-spectrum binding and inhibition, and we devised a multiplexed poly-target selection to elicit those phenotypes against a panel of primate lentiviral RTs. High-throughput sequencing and coenrichment/codepletion analysis of parallel and duplicate selection trajectories rapidly narrowed the list of candidate aptamers by orders of magnitude and identified dozens of priority candidates for further screening. Biochemical characterization validated a novel aptamer motif and several rare and unobserved variants of previously known motifs that inhibited recombinant RTs to varying degrees. These broad-spectrum aptamers also suppressed replication of viral constructs carrying phylogenetically diverse RTs. The poly-target selection and coenrichment/codepletion approach described herein is a generalizable strategy for identifying cross-reactivity among related targets from combinatorial libraries. Graphical Abstract |
| Databáze: | OpenAIRE |
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