Improved Delivery in Cell Culture of Radiolabeled Antisense DNAs by Duplex Formation
Improved Delivery in Cell Culture of Radiolabeled Antisense DNAs by Duplex Formation
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DOI：
10.1007/s11307-006-0050-7
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发表时间：
2006-08
影响因子：
3.1
通讯作者：
Xinrong Liu;Kayoko Nakamura;Yi Wang;Surong Zhang;Jiang He;Guozheng Liu;S. Dou;A. Kubo;M. Rusckowski;D. Hnatowich
中科院分区：
文献类型：
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作者：
Xinrong Liu;Kayoko Nakamura;Yi Wang;Surong Zhang;Jiang He;Guozheng Liu;S. Dou;A. Kubo;M. Rusckowski;D. Hnatowich
目的在需要静脉注射 DNA 的应用中，递送仍然是一个未解决的问题。据报道，最近，与单线态 AS 寡聚物相比，双链体反义 (AS) 寡聚物在细胞中的反义翻译中断得到了改善，这可能是因为递送得到了改善。有义 (S) 寡聚物不稳定的磷酸二酯主链及其较短的链长显然促进了 AS 寡聚物的细胞内解离和释放。我们研究了所涉及的机制，以评估该方法是否可用于反义放射性核素成像。程序双链体是在针对themdr1mRNA的AS硫代磷酸酯DNA和具有四个或六个错配的统一硫代磷酸酯或统一磷酸二酯正义（S）DNA之间形成的。结果放射性标记的AS DNA在KB-G2（Pgp++）细胞中的累积量作为双链体累积量比单链体高三倍。如 S DNA 上放射性标记的积累减少所示，积累仍然是反义的。然而，DNA 主链对积累没有明显的影响。结论如果与设计为低杂交亲和力的 S DNA 双链，以在目标 mRNA 存在的情况下促进解离，则在细胞培养物中用放射性标记的 AS DNA 靶向 mRNA 可能会得到改善。
PurposeDelivery remains an unresolved problem in applications requiring intravenous administration of DNAs. Recently improved antisense translation interruption in cells was reported for an antisense (AS) oligomer as a duplex compared to singlet AS oligomer presumably because of improved delivery. The unstable phosphodiester backbone of the sense (S) oligomer and its shorter chain length apparently encouraged intracellular dissociation and release of the AS oligomer. We have investigated the mechanism involved to evaluate whether the approach may be useful for antisense radionuclide imaging.ProceduresDuplexes were formed between an AS phosphorothioate DNA against themdr1mRNA and the uniform phoshorothioate or uniform phosphodiester sense (S) DNAs with either four or six mismatches.ResultsAccumulations in KB-G2 (Pgp++) cells of radiolabeled AS DNA as duplex accumulated threefold higher compared to singlet. Accumulation was still antisense as shown by reduced accumulations with the radiolabel on the S DNA. However, the DNA backbone had no clear influence on accumulations.ConclusionsTargeting of mRNAs with radiolabeled AS DNAs may be improved in cell culture if duplexed with an S DNA engineered for low hybridization affinity to encourage dissociation in the presence of the target mRNA.