2'-fluoro-modified pyrimidines enhance affinity of RNA oligonucleotides to HIV-1 reverse transcriptase.
2'-fluoro-modified pyrimidines enhance affinity of RNA oligonucleotides to HIV-1 reverse transcriptase.
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DOI：
10.1261/rna.077008.120
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发表时间：
2020-11
期刊：
影响因子：
--
通讯作者：
Burke DH
中科院分区：
文献类型：
--
作者：
Gruenke PR;Alam KK;Singh K;Burke DH
核酸适体可以被化学修饰以增强功能，但是修饰先前选择的适体可以具有重要的结构和功能后果。我们提出了一种重新选择策略，以评估几个修改对预先存在的适体池的影响。用2′-F、2′-OMe或2′-NH 2嘧啶逆转录对HIV-1逆转录酶（RT）具有亲和力的RNA适体文库，并进行三个额外的选择循环。RT抑制被观察到的代表性适体从几个结构的家庭确定高通量测序时，转录与其相应的修改。因此，重选鉴定了耐受来自未修饰的预富集文库的化学修饰的适体的专门子集。与2′-OMeY和2′-NH 2 Y RNA的抑制相比，2′-F-嘧啶（2′-FY）RNA的抑制作用最强。出乎意料的是，一组不同的逆转录病毒RT被所有2′-FY修饰的转录物强烈抑制，包括不抑制这些RT的序列作为未修饰的RNA。混杂RT抑制的大小与转录物中2′-FY的摩尔分数成正比。2′-FY转录物的RT结合亲和力对盐浓度比未修饰转录物的结合更敏感，表明2′-FY RNA与逆转录病毒RT的相互作用比未修饰的2′-OH RNA更具离子性。2′-FY修饰RNA的这些令人惊讶的特征可能对应用适体技术具有普遍意义。
Nucleic acid aptamers can be chemically modified to enhance function, but modifying previously selected aptamers can have nontrivial structural and functional consequences. We present a reselection strategy to evaluate the impact of several modifications on preexisting aptamer pools. RNA aptamer libraries with affinity to HIV-1 reverse transcriptase (RT) were retranscribed with 2′-F, 2′-OMe, or 2′-NH2 pyrimidines and subjected to three additional selection cycles. RT inhibition was observed for representative aptamers from several structural families identified by high-throughput sequencing when transcribed with their corresponding modifications. Thus, reselection identified specialized subsets of aptamers that tolerated chemical modifications from unmodified preenriched libraries. Inhibition was the strongest with the 2′-F-pyrimidine (2′-FY) RNAs, as compared to inhibition by the 2′-OMeY and 2′-NH2Y RNAs. Unexpectedly, a diverse panel of retroviral RTs were strongly inhibited by all 2′-FY-modified transcripts, including sequences that do not inhibit those RTs as unmodified RNA. The magnitude of promiscuous RT inhibition was proportional to mole fraction 2′-FY in the transcript. RT binding affinity by 2′-FY transcripts was more sensitive to salt concentration than binding by unmodified transcripts, indicating that interaction with retroviral RTs is more ionic in character for 2′-FY RNA than for unmodified 2′-OH RNA. These surprising features of 2′-FY-modified RNA may have general implications for applied aptamer technologies.
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影响因子：
14.9
作者：
AURUP, H;TUSCHL, T;ECKSTEIN, F
通讯作者：
ECKSTEIN, F
影响因子：
12.4
作者：
Lange, Margaret J.;Sharma, Tarun K.;Burke, Donald H.
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Burke, Donald H.
影响因子：
5.6
作者：
GREEN, L;WAUGH, S;TUERK, C
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TUERK, C
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1.6
作者：
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通讯作者：
Inouye, Kuniyo
影响因子：
--
作者：
Burnett JC;Rossi JJ
通讯作者：
Rossi JJ