Nucleic Acids Research, ( ISI ), Volume (38), No (3), Year (2010-1) , Pages (1048-1056)

Title : ( A single-label phenylpyrrolocytidine provides a molecular beacon-like response reporting HIV-1 RT RNase H activity )

Authors: Alexander S. Wahba , Abbas Ali Esmaeili , Masad J. Damha , Robert H. E. Hudson ,

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6-Phenylpyrrolocytidine (PhpC), a structurally conservative and highly fluorescent cytidine analog, was incorporated into oligoribonucleotides. The PhpC-containing RNA formed native-like duplex structures with complementary DNA or RNA. The PhpC-modification was found to act as a sensitive reporter group being non-disruptive to structure and the enzymatic activity of RNase H. A RNA/DNA hybrid possessing a single PhpC insert was an excellent substrate for HIV-1 RT Ribonuclease H and rapidly reported cleavage of the RNA strand with a 14-fold increase in fluorescence intensity. The PhpC-based assay for RNase H was superior to the traditional molecular beacon approach in terms of responsiveness, rapidity and ease (single label versus dual). Furthermore, the PhpC-based assay is amenable to high-throughput microplate assay format and may form the basis for a new screen for inhibitors of HIV-RT RNase H.

Keywords

, phenylpyrrolocytidine, HIV-1 RT RNase H
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@article{paperid:1025925,
author = {Alexander S. Wahba and Esmaeili, Abbas Ali and Masad J. Damha and Robert H. E. Hudson},
title = {A single-label phenylpyrrolocytidine provides a molecular beacon-like response reporting HIV-1 RT RNase H activity},
journal = {Nucleic Acids Research},
year = {2010},
volume = {38},
number = {3},
month = {January},
issn = {0305-1048},
pages = {1048--1056},
numpages = {8},
keywords = {phenylpyrrolocytidine; HIV-1 RT RNase H activity},
}

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%0 Journal Article
%T A single-label phenylpyrrolocytidine provides a molecular beacon-like response reporting HIV-1 RT RNase H activity
%A Alexander S. Wahba
%A Esmaeili, Abbas Ali
%A Masad J. Damha
%A Robert H. E. Hudson
%J Nucleic Acids Research
%@ 0305-1048
%D 2010

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