固相合成,热变性研究,核酶抵抗,以及细胞吸收 (oligodeoxyribonucleoside) 甲基酸-DNA仿真体
Heera Krishna1, Marvin H Caruthers
1Department of Chemistry and Biochemistry, University of Colorado at Boulder, Boulder, Colorado 80309, USA.
Journal of the American Chemical Society
|May 19, 2011
概括
新的甲基酸 (Me-P-BH(3)) oligodeoxynucleotides (ODNs) 显示出高的核酶抵抗性和高效的细胞吸收. 这些修改后的ODN对反意义研究和RNA干扰 (RNAi) 应用具有前景.
科学领域:
- 药用化学 医学化学
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 氧化氧核酸 (ODN) 面临治疗用途的挑战,原因是细胞传递不良和核酶降解.
- 甲基酸提供核酶稳定性,而酸提供良好的膜透性和基因沉默.
- 一个新的修改,甲基酸 (Me-P-BH(3)),旨在结合两者的优势.
研究的目的:
- 合成和表征混合脊柱寡度氧核酸 (ODN) 结合甲基酸 (Me-P-BH(3)) 链接.
- 评估这些新型ODN的核酶稳定性,杂交特性和细胞吸收.
- 评估Me-P-BH(3) ODNs在反意义和RNA干扰 (RNAi) 研究中的潜力.
主要方法:
- 自动固体相合成16-mer混合骨干ODN与甲基酸和酸/酸链接.
- 核酶稳定性测试用于测试对5'和3'外核酶的抗性.
- 杂交研究以形成ODN:RNA复杂体并评估结合亲和力.
- 在HeLa和WM-239A细胞中使用光标记的ODN进行细胞吸收研究.
主要成果:
- 甲基酸ODNs表现出对5'和3'外核酶的高耐药性.
- 与原生DNA:RNA复合体相比,ODN:RNA复合体表现出增强的稳定性,在较低的盐度下,结合亲和力增加.
- 观察到 Me-P-BH 的有效细胞吸收,3 ODN,治疗细胞中显著的细胞质光.
- 似乎Me-P-BH(3) 链接具有正电荷,有助于改善结合和吸收.
结论:
- 甲基酸 (Me-P-BH(3)) ODNs在核酶稳定性和细胞传递方面提供了显著的改进.
- 这些修改后的ODN与RNA形成稳定的重复体,这表明基因沉默应用的潜力.
- (3) ODN代表了一个有前途的新分子类别,用于反意义和RNAi策略的治疗开发.
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