Medicinal Chemistry, Rega Institute for Medical Research, KU Leuven, Herestraat 49, Box-1041, 3000, Leuven, Belgium.
Angew Chem Int Ed Engl. 2021 Feb 19;60(8):4175-4182. doi: 10.1002/anie.202011887. Epub 2020 Dec 27.
Synthetic nucleic acids, with four non-canonical nucleobases, can function as genetic materials. A comprehensive analysis of PCR amplification, transcription, reverse transcription, and cloning was done to screen for alternative genetic monomers. A small library of six modified nucleobases was selected: the modified 2'-deoxyribonucleoside (dZTPs) and ribonucleoside (rZTPs) triphosphates of 7-deaza-adenine, 5-chlorouracil, 7-deaza-guanine or inosine together with 5-fluorocytosine or 5-bromocytosine. The fragments composed of one to four modified nucleotides (denoted as DZA) have been successfully recognized and transcribed to natural or modified RNA (denoted as RZA) by T7 RNA polymerase. The fully modified RZA fragment could be reverse transcribed and then amplified in the presence of various dZTPs. Noticeably, modified fragments could function as genetic templates in vivo by encoding the 678 base pair gene of a fluorescent protein in bacteria. These results demonstrate the existence of a fully simulated genetic circuit that uses synthetic materials.
合成核酸，具有四个非经典碱基，可用作遗传物质。我们全面分析了聚合酶链反应扩增、转录、反转录和克隆，以筛选替代遗传单体。我们选择了一个包含六种修饰碱基的小文库：7-脱氮腺嘌呤、5-氯尿嘧啶、7-脱氮鸟嘌呤或肌苷的修饰 2'-脱氧核苷（dZTPs）和核糖核苷（rZTPs）三磷酸与 5-氟胞嘧啶或 5-溴胞嘧啶一起。由一个到四个修饰核苷酸组成的片段（表示为 DZA）已被 T7 RNA 聚合酶成功识别并转录为天然或修饰的 RNA（表示为 RZA）。完全修饰的 RZA 片段可以在存在各种 dZTP 的情况下反转录和扩增。值得注意的是，修饰片段可以在体内作为遗传模板，在细菌中编码荧光蛋白的 678 个碱基对基因。这些结果表明，使用合成材料存在一个完全模拟的遗传电路。
