An efficient unnatural base pair for a base-pair-expanded transcription system
An efficient unnatural base pair for a base-pair-expanded transcription system
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DOI：
10.1021/ja0425280
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发表时间：
2005-06-22
影响因子：
15
通讯作者：
Hirao, L
中科院分区：
文献类型：
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作者：
Mitsui, T;Kimoto, M;Hirao, L
为了通过转录将人工成分定位到RNA中，开发了2-氨基-6-(2-噻唑基)嘌呤(记为v)和2-氧(1H)吡啶(记为y)之间的高效非自然碱基对。通过模板中与v相反的T7RNA聚合酶将y和5-取代y的底物特异性地掺入到RNA中。在转录过程中，v-y配对的效率和保真度与自然A-T(U)和G-C配对一样高。此外，还从含有两个v碱基的DNA模板转录出含有两个相邻y碱基的RNA。这种特定的转录允许大规模制备人造RNA，并可以与其他系统结合，同时将几个不同的成分合并到转录本中。
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.