扩大可复制的非自然DNA的范围:逐步优化一种主要是疏水的基对
Thomas Lavergne1, Mélissa Degardin, Denis A Malyshev
1Department of Chemistry and Center for Protein and Nucleic Acid Research, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
Journal of the American Chemical Society
|April 4, 2013
概括
研究人员通过创造新的疏水性非自然基因对来扩展遗传字母. 最有前途的衍生品d5SICS-dFEMO显示了增强的复制效率和真实性,为体内应用铺平了道路.
科学领域:
- 合成生物学和基因工程学
- 核酸化学和分子生物学分子生物学
背景情况:
- 目前正在努力扩大用于体外和体内生物应用的遗传字母.
- 之前的研究已经确定d5SICS-dMMO2和d5SICS-dNaM是疏水性非自然基对.
- 优化d5SICS-dNaM突出了dMMO2的具体位置,以便进一步开发.
研究的目的:
- 通过衍生来系统地优化dMMO2基对.
- 评估para和meta替代剂对复制和转录效率和真实性的影响.
- 为了确定新的非自然基对,以提高潜在的 in vivo 应用的性能.
主要方法:
- 合成和评估dMMO2.2的18种新型副衍生类型的类似物.
- 进一步衍生和评估有前途的类似物与元替代物.
- 评估DNA内修改基因对的复制和转录效率和忠实性.
主要成果:
- 副替代剂可以通过硬质和电子效应优化DNA复制.
- 甲基甲基组被发现是不利的,而替代剂显示了可变的效果.
- 不自然三酸盐插入效率的改善与更高的复制保真度相关.
- 包括d5SICS-dFEMO在内的多种新基对衍生物证明了有效和准确的复制.
- 在某些条件下,d5SICS-dFEMO表现出比d5SICS-dNaM更高或可比的效率和保真度.
结论:
- 疏水力通常适用于控制DNA内的基配对.
- 广泛的结构-活性关系 (SAR) 数据被生成用于非自然的基对优化.
- 一些新的非自然基对,特别是d5SICS-dFEMO,是未来体内研究的有希望的候选者.
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