Towards mRNA with superior translational activity: synthesis and properties of ARCA tetraphosphates with single phosphorothioate modifications.
Towards mRNA with superior translational activity: synthesis and properties of ARCA tetraphosphates with single phosphorothioate modifications.
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DOI：
10.1039/b9nj00644c
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发表时间：
2010-01-01
期刊：
影响因子：
--
通讯作者：
Jemielity J
中科院分区：
文献类型：
--
作者：
Strenkowska M;Kowalska J;Lukaszewicz M;Zuberek J;Su W;Rhoads RE;Darzynkiewicz E;Jemielity J
我们描述了一系列mRNA 5'端（帽）类似物的化学合成和初步的生物物理和生物化学表征，这些类似物被设计为用于获得具有增强的翻译效率和体内稳定性的mRNA分子的试剂，以及作为研究mRNA代谢的有用工具。类似物共有三个结构特征：（i）5 '，5'-桥延长为四磷酸以增加它们对翻译起始因子eIF 4 E的亲和力（ii）在α，β，四磷酸的γ或δ-位，以降低它们对酶促降解的敏感性和/或调节它们与特定蛋白质的相互作用，以及（iii）2 '-O-7-甲基鸟苷核糖中的甲基，抗反向帽类似物（ARCA）的特征，在体外转录过程中仅以正确的方向掺入mRNA中。通过ZnCl_2介导的两个单核苷酸亚基之间的偶联合成了具有修饰的四磷酸桥的二核苷酸，分离产率为30- 65%。初步的生物化学结果表明，用新类似物封端的mRNA在无细胞系统中的翻译效率比m7 GpppG封端的mRNA高2.5-4.5倍，这使得它们成为基于RNA的治疗应用如基因治疗和抗癌疫苗的有希望的候选者。
We describe the chemical synthesis and preliminary biophysical and biochemical characterization of a series of mRNA 5' end (cap) analogs designed as reagents for obtaining mRNA molecules with augmented translation efficiency and stability in vivo and as useful tools to study mRNA metabolism. The analogs share three structural features: (i) 5',5'- bridge elongated to tetraphosphate to increase their affinity to translation initiation factor eIF4E (ii) a single phosphorothioate modification at either the α, β, γ or δ-position of the tetraphosphate to decrease their susceptibility to enzymatic degradation and/or to modulate their interaction with specific proteins and (iii) a 2'-O-methyl group in the ribose of 7-methylguanosine, characteristic to Anti-Reverse Cap Analogs (ARCAs), which are incorporated into mRNA during in vitro transcription exclusively in the correct orientation. The dinucleotides bearing modified tetraphosphate bridge were synthesized by ZnCl2 mediated coupling between two mononucleotide subunits with isolated yields of 30–65%. The preliminary biochemical results show that mRNAs capped with new analogs are 2.5–4.5 more efficiently translated in a cell free system than m7GpppG-capped mRNAs, which makes them promising candidates for RNA-based therapeutic applications such as gene therapy and anti-cancer vaccines.
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影响因子：
12.4
作者：
Kariko, Katalin;Muramatsu, Hiromi;Welsh, Frank A.;Ludwig, Janos;Kato, Hiroki;Akira, Shizuo;Weissman, Drew
通讯作者：
Weissman, Drew
影响因子：
4.5
作者：
Liu, SW;Jiao, XF;Kiledjian, M
通讯作者：
Kiledjian, M
影响因子：
4.8
作者：
Grudzien, E;Kalek, M;Rhoads, RE
通讯作者：
Rhoads, RE
影响因子：
20.3
作者：
Holtkamp, Silke;Kreiter, Sebastian;Sahin, Ugur
通讯作者：
Sahin, Ugur
影响因子：
4.5
作者：
Grudzien-Nogalska, Ewa;Jemielity, Jacek;Rhoads, Robert E.
通讯作者：
Rhoads, Robert E.