KU Leuven, Rega Institute for Medical Research, Medicinal Chemistry, Herestraat 49, Box 1041, 3000, Leuven, Belgium.
School of Biology and Environmental Science, Queensland University of Technology, Brisbane, QLD, 4001, Australia.
Chembiochem. 2022 Jun 3;23(11):e202200060. doi: 10.1002/cbic.202200060. Epub 2022 Apr 12.
Chemically modified nucleic acids are of utmost interest in synthetic biology for creating a regulable and sophisticated synthetic system with tailor-made properties. Implanting chemically modified nucleic acids in microorganisms might serve biotechnological applications, while using them in human cells might lead to new advanced medicines. Previously, we reported that a fully modified DNA sequence (called DZA) composed of the four base-modified nucleotides - 7-deaza-adenine, 5-chlorouracil, 7-deaza-guanine and 5-fluorocytosine - could function as a genetic template in prokaryotic cells, Escherichia coli. Here, we report the synthesis of long, partially, or fully modified DZA fragments that encode the yeast-enhanced red fluorescent protein (yEmRFP). The DZA sequences were directly introduced in the genome of the eukaryotic cells, Saccharomyces cerevisiae, via the yeast natural homologous recombination machinery. The simple and straightforward DZA cloning strategy reported here might be of interest to scientists working in the field of xenobiology in yeast.
化学修饰的核酸在合成生物学中具有重要意义，可用于构建具有定制特性的可调控和复杂的合成系统。将化学修饰的核酸植入微生物中可能用于生物技术应用，而将其用于人体细胞可能会产生新的高级药物。此前，我们曾报道过一个完全由碱基修饰核苷酸组成的全修饰 DNA 序列（称为 DZA），该序列由 7-脱氮腺嘌呤、5-氯尿嘧啶、7-脱氮鸟嘌呤和 5-氟胞嘧啶四种碱基修饰核苷酸组成，可在原核细胞大肠杆菌中作为遗传模板起作用。在此，我们报告了长链、部分或全修饰 DZA 片段的合成，这些片段编码酵母增强型红色荧光蛋白（yEmRFP）。通过酵母天然同源重组机制，DZA 序列可直接被引入真核细胞酿酒酵母的基因组中。这里报道的简单直接的 DZA 克隆策略可能会引起从事酵母外来生物领域研究的科学家的兴趣。
