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使用胺酸化学合成胺酸和它们的奇米拉

Sibasish Paul¹, Marvin H Caruthers¹

¹Department of Chemistry and Biochemistry, University of Colorado , Boulder, Colorado 80309, United States.

Journal of the American Chemical Society

|December 10, 2016

概括

合成了新的二胺酸 (PMO) 模拟物和PMO-DNA仿真体. 一些修改后的PMO具有增强的结合和刺激RNaseH1活性,从而促进了寡核酸的研究.

科学领域:

氧核酸化学
分子生物学
生物技术

背景情况:

二胺酸 (PMOs) 是合成的核酸类似物.
在反意义技术,诊断和纳米技术中使用PMO.
开发具有改进性质的新型PMO类似物对于扩大其应用至关重要.

研究的目的:

合成新的二胺酸 (PMO) 类似物和PMO-DNA模拟物.
评估这些改性寡核酸的结合亲和力和稳定性.
研究PMO类型的生物活性,包括RNaseH1刺激和细胞吸收.

主要方法:

使用胺化工合成PMO和PMO-DNA仿真体.
酸胺酸核酸间链接的制备.
用各种氨基 (N,N-二甲基胺,N-甲基胺,氨,形态素) 进行氧化替代.
用DNA和RNA对复合体进行化温度 (Tm) 分析.
评估RNaseH1活动的刺激.
使用光标记的PMO仿真体在用脂质转染试剂的HeLa细胞中的细胞吸收研究.

主要成果:

与天然的DNA复合体相比,氨基基改性PMO具有更高的化温度.
具有Morpholino,Dimethylamino或N-methylamino结合的PMO类似物显示出可比或降低的化温度.
N,N-二甲基氨基PMO-DNA仿真体刺激了RNaseH1的活性.
光标记的PMO仿真体在使用脂质转染试剂时,在HeLa细胞中表现出高效的细胞吸收.

结论:

简化合成使得PMO类似物容易获得.
修改后的PMO为寡核酸应用提供可调节的特性.
这些发现为反意义,诊断和纳米技术寡核酸领域的研究开辟了新的途径.

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