Department of Advanced Science and Technology, Japan Advanced Institute of Science and Technology, 1-1 Asahidai, Nomi, Ishikawa 923-1211, Japan.
Molecules. 2018 Apr 4;23(4):828. doi: 10.3390/molecules23040828.
Gene therapies, including genome editing, RNAi, anti-sense technology and chemical DNA editing are becoming major methods for the treatment of genetic disorders. Techniques like CRISPR-Cas9, zinc finger nuclease (ZFN) and transcription activator-like effector-based nuclease (TALEN) are a few such enzymatic techniques. Most enzymatic genome editing techniques have their disadvantages. Thus, non-enzymatic and non-invasive technologies for nucleic acid editing has been reported in this study which might possess some advantages over the older methods of DNA manipulation. 3-cyanovinyl carbazole (K) based nucleic acid editing takes advantage of photo-cross-linking between a target pyrimidine and the K to afford deamination of cytosine and convert it to uracil. This method previously required the use of high temperatures but, in this study, it has been optimized to take place at physiological conditions. Different counter bases (inosine, guanine and cytosine) complementary to the target cytosine were used, along with derivatives of K (K and K) to afford the deamination at physiological conditions.
基因疗法，包括基因组编辑、RNAi、反义技术和化学 DNA 编辑，正在成为治疗遗传疾病的主要方法。CRISPR-Cas9、锌指核酸酶 (ZFN) 和转录激活因子样效应物核酸酶 (TALEN) 等酶技术就是其中的几种。大多数酶促基因组编辑技术都有其缺点。因此，本研究报道了非酶促和非侵入性的核酸编辑技术，它可能比旧的 DNA 操作方法具有一些优势。基于 3-氰基乙烯基咔唑 (K) 的核酸编辑利用靶嘧啶与 K 之间的光交联，使胞嘧啶脱氨并转化为尿嘧啶。该方法以前需要使用高温，但在本研究中，它已被优化为在生理条件下进行。使用了与靶胞嘧啶互补的不同的反碱基（次黄嘌呤、鸟嘌呤和胞嘧啶），以及 K（K 和 K）的衍生物，以在生理条件下进行脱氨。
