Driving In Vitro Selection of Anti-HIV-1 TAR Aptamers by Magnesium Concentration and Temperature

Authors: Darfeuille F.; Sekkai D.; Dausse E.; Kolb G.; Yurchenko L.; Boiziau C.; Toulme J.J.

Source: Combinatorial Chemistry & High Throughput Screening, Volume 5, Number 4, June 2002 , pp. 313-325(13)

Publisher: Bentham Science Publishers

Abstract:

In vitro selection with either DNA or RNA libraries was performed against the TAR RNA element of HIV-1. The role of the selection conditions on the outcome of the selection was evaluated by varying the magnesium concentration and the temperature. The selection stringency was demonstrated to determine i) the affinity of the best identified aptamers for the TAR target, and ii) the type of interaction between the two partners. Selections performed with a DNA library under low (4 C, 10 mM magnesium) and high stringency (23 C, 3 mM magnesium) led to the emergence of “kissing aptamers” but even if the motif interacting directly with the TAR loop were identical in the two kinds of aptamers, the consensus was extended from eight to thirteen nucleotides when the Mg2+ concentration was decreased from 10 to 3 mM. Similar kissing aptamers were selected at 23 C and 37 C starting with two different RNA libraries under identical ionic conditions. In addition, selection performed at 37 C yielded a significant proportion of antisense sequences. Only antisense RNAs complementary to the TAR loop competitively inhibited the association of a Tat peptide with TAR.

Keywords: Anti-HIV-1 TAR Aptamers; Oligonucleotides; TAR Loop; Tat Binding

Language: English

Document Type: Review article

DOI: http://dx.doi.org/10.2174/1386207023330282

Publication date: 2002-06-01

• Combinatorial Chemistry & High Throughput Screening publishes full length original research articles and reviews describing various topics in combinatorial chemistry (e.g. small molecules, peptide, nucleic acid or phage display libraries) and/or high throughput screening (e.g. developmental, practical or theoretical). Ancillary subjects of key importance, such as robotics and informatics, will also be covered by the journal. In these respective subject areas, Combinatorial Chemistry & High Throughput Screening is intended to function as the most comprehensive and up-to-date medium available. The journal should be of value to individuals engaged in the process of drug discoveryand development, in the settings of industry, academia or government.

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• By this author: Darfeuille F. ;  Sekkai D. ;  Dausse E. ;  Kolb G. ;  Yurchenko L. ;  Boiziau C. ;  Toulme J.J.

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