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21.
Pavel A. Sakharov Egor A. Smolin Dmitry N. Lyabin Sultan C. Agalarov 《International journal of molecular sciences》2021,22(7)
The methylation of adenosine in the N6 position (m6A) is a widely used modification of eukaryotic mRNAs. Its importance for the regulation of mRNA translation was put forward recently, essentially due to the ability of methylated mRNA to be translated in conditions of inhibited cap-dependent translation initiation, e.g., under stress. However, the peculiarities of translation initiation on m6A-modified mRNAs are not fully known. In this study, we used toeprinting and translation in a cell-free system to confirm that m6A-modified mRNAs can be translated in conditions of suppressed cap-dependent translation. We show for the first time that m6A-modified mRNAs display not only decreased elongation, but also a lower efficiency of translation initiation. Additionally, we report relative resistance of m6A-mRNA translation initiation in the absence of ATP and inhibited eIF4A activity. Our novel findings indicate that the scanning of m6A-modified leader sequences is performed by a noncanonical mechanism. 相似文献
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Tomasz Z. Tomkiewicz Nuria Surez-Herrera Frans P. M. Cremers Rob W. J. Collin Alejandro Garanto 《International journal of molecular sciences》2021,22(9)
The discovery of novel intronic variants in the ABCA4 locus has contributed significantly to solving the missing heritability in Stargardt disease (STGD1). The increasing number of variants affecting pre-mRNA splicing makes ABCA4 a suitable candidate for antisense oligonucleotide (AON)-based splicing modulation therapies. In this study, AON-based splicing modulation was assessed for 15 recently described intronic variants (three near-exon and 12 deep-intronic variants). In total, 26 AONs were designed and tested in vitro using a midigene-based splice system. Overall, partial or complete splicing correction was observed for two variants causing exon elongation and all variants causing pseudoexon inclusion. Together, our results confirm the high potential of AONs for the development of future RNA therapies to correct splicing defects causing STGD1. 相似文献
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Jinguo Huang Ruiqi Zhao Jing Mo Fang Wang Dr. Xiaocheng Weng Xiang Zhou 《Chembiochem : a European journal of chemical biology》2021,22(9):1559-1562
There is growing interest in developing intracellular RNA tools. Herein, we describe a strategy for N3-kethoxal (N3K)-based bioorthogonal intracellular RNA functionalization. With N3K labeling followed by an in vivo click reaction with DBCO derivatives, RNA can be modified with fluorescent or phenol groups. This strategy provides a new way of labeling RNA inside cells. 相似文献
27.
Renner S Ludwig V Boden O Scheffer U Göbel M Schneider G 《Chembiochem : a European journal of chemical biology》2005,6(6):1119-1125
TAR RNA is a potential target for AIDS therapy. Ligand-based virtual screening was performed to retrieve novel scaffolds for RNA-binding molecules capable of inhibiting the Tat-TAR interaction, which is essential for HIV replication. We used a "fuzzy" pharmacophore approach (SQUID) and an alignment-free pharmacophore method (CATS3D) to carry out virtual screening of a vendor database of small molecules and to perform "scaffold-hopping". A small subset of 19 candidate molecules were experimentally tested for TAR RNA binding in a fluorescence resonance energy transfer (FRET) assay. Both methods retrieved molecules that exhibited activities comparable to those of the reference molecules acetylpromazine and chlorpromazine, with the best molecule showing ten times better binding behavior (IC50 = 46 microM). The hits had molecular scaffolds different from those of the reference molecules. 相似文献
28.
Verhelst SH Michiels PJ van der Marel GA van Boeckel CA van Boom JH 《Chembiochem : a European journal of chemical biology》2004,5(7):937-942
Aminoglycoside antibiotics, which are able to selectively bind to RNA, are considered to be an important lead in RNA-targeting drug discovery. In this study, surface plasmon resonance (SPR) was employed to explore the interaction of aminoglycosides with known tobramycin-binding RNA hairpins (aptamers) and an unrelated RNA hairpin. It was established that aminoglycosides have multiple interactions with RNA hairpins. Unexpectedly, the different hairpins showed comparable affinity for a set of related aminoglycosides. The observed absence of selectivity presents an extra hurdle in the discovery of novel aminoglycosides as specific drugs that target defined RNA hairpins. 相似文献
29.
几家HCV抗体诊断试剂盒检测系列血清的比较 总被引:3,自引:0,他引:3
应用Abbott、UBI及国内几家的HCV第二代抗体诊断试剂盒A、B、c及D检测25名HCV感染者的210份感染后不同时期的系列血清,首次采用ELISA及RIBA分片段检测抗体,并与敏感的PCR法检测血清中HCV RNA结果相比较。虽然各试剂盒的总体检测符合率无显著性差异,但个别国产试剂盒的早期检测能力尚有待提高。用任何一家抗体检测试剂盒均会漏检小部分标本,抗体检测阴性的血清在100000倍稀释后仍可检出HCV RNA,因此,有必要发展包括更多片段包被的第三代HCV抗体诊断试剂盒。 相似文献
30.
针对甲型流感病毒H1N1基因,从RNAi的角度出发,采用多特征融合的方法,进行siRNA预测。对2009年的46株病毒序列的PA片段进行分析,从经过序列分析所获得的众多靶序列中,采用结构分析手段对靶序列进行筛选,获得较易干扰的靶序列及设计出相应的siRNA。2009年爆发的H1N1病毒,序列保守性高,靶序列一致性高,结构保守性高。该方法可以有效选择可能的靶序列,并在此基础上进一步筛选,以获得少量较易干扰的靶序列,该方法为复杂序列siRNA的设计提供了新思路,对siRNA的优化设计有指导意义,有助于利用RNAi进行H1N1治疗的后续研究。 相似文献