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RNA G-Quadruplex Invasion and Translation Inhibition by Antisense gamma-Peptide Nucleic Acid Oligomers

机译:反义γ-肽核酸寡聚物对RNA G-四链体的侵袭和翻译抑制

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摘要

We have examined the abilities of three complementary gamma-peptide nucleic acid (gamma PNA) oligomers to invade an RNA G-quadruplex and potently inhibit translation of a luciferase reporter transcript containing the quadruplex-forming sequence (QFS) within its 5'-untranslated region. All three gamma PNA oligomers bind with low nanomolar affinities to an RNA oligonucleotide containing the QFS. However, while all probes inhibit translation with low to midnanomolar IC50 values, the gamma PNA designed to hybridize to the first two G-tracts of the QFS and adjacent 5'-overhanging nucleotides was 5-6 times more potent than probes directed to either the 3'-end or internal regions of the target at 37 degrees C. This position-dependent effect was eliminated after the probes and target were preincubated at an elevated temperature prior to translation, demonstrating that kinetic effects exert significant control over quadruplex invasion and translation inhibition. We also found that antisense gamma PNAs exhibited similarly potent effects against luciferase reporter transcripts bearing QFS motifs having G(2), G(3), or G(4) tracts. Finally, our results indicate that gamma PNA,oligomers exhibit selectivity and/or potency higher than those of other antisense molecules such as standard PNA and 2'-OMe RNA previously reported to target G-quadruplexes in RNA.
机译:我们研究了三种互补的 γ-肽核酸 (gamma PNA) 寡聚体侵入 RNA G-四链体并有效抑制在其 5'-非翻译区域内包含四链体形成序列 (QFS) 的荧光素酶报告基因转录本的翻译的能力。所有三种γ PNA寡聚物都以低纳摩尔亲和力与含有QFS的RNA寡核苷酸结合。然而,虽然所有探针都以低至中纳摩尔 IC50 值抑制翻译,但设计用于与 QFS 的前两个 G 束和相邻的 5'-悬垂核苷酸杂交的 γ PNA 比在 37 摄氏度时指向靶标的 3' 末端或内部区域的探针强 5-6 倍。在翻译前将探针和靶标在高温下预孵育后,这种位置依赖性效应被消除,表明动力学效应对四链体侵袭和平移抑制具有显着的控制作用。我们还发现,反义γ PNA对带有QFS基序的荧光素酶报告基因转录本表现出类似的有效作用,这些基序具有G(2)、G(3)或G(4)束。最后,我们的结果表明,γ PNA,寡聚物表现出高于其他反义分子的选择性和/或效力,例如标准PNA和先前报道的靶向RNA中的G-四链体的2'-OMe RNA。

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