一种用于碱基对扩展转录系统的高效非天然碱基对。

An efficient unnatural base pair for a base-pair-expanded transcription system.

作者信息

Mitsui Tsuneo, Kimoto Michiko, Harada Yoko, Yokoyama Shigeyuki, Hirao Ichiro

机构信息

Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.

出版信息

J Am Chem Soc. 2005 Jun 22;127(24):8652-8. doi: 10.1021/ja0425280.

DOI:10.1021/ja0425280

PMID:15954770

Abstract

For the site-specific incorporation of artificial components into RNA by transcription, an efficient, unnatural base pair between 2-amino-6-(2-thiazolyl)purine (denoted as v) and 2-oxo(1H)pyridine (denoted as y) was developed. The substrates of y and 5-substituted y were site-specifically incorporated into RNA by T7 RNA polymerase opposite v in templates. The efficiency and fidelity of the v-y pairing in transcription were as high as those of the natural A-T(U) and G-C pairings. Furthermore, RNAs containing two adjacent y bases were also transcribed from DNA templates containing two v bases. This specific transcription allows the large-scale preparation of artificial RNAs and can be combined with other systems to simultaneously incorporate several different components into a transcript.

摘要

为了通过转录将人工成分位点特异性地掺入RNA中，人们开发了一种在2-氨基-6-（2-噻唑基）嘌呤（表示为v）和2-氧代（1H）吡啶（表示为y）之间的高效非天然碱基对。y和5-取代的y底物通过T7 RNA聚合酶在模板中与v相对应位点特异性地掺入RNA中。转录过程中v-y配对的效率和保真度与天然A-T（U）和G-C配对一样高。此外，含有两个相邻y碱基的RNA也从含有两个v碱基的DNA模板转录而来。这种特异性转录允许大规模制备人工RNA，并且可以与其他系统结合，将几种不同的成分同时掺入转录本中。