College of Inter-Faculty Individual Studies in Mathematics and Natural Sciences, Banacha 2c, 02-097, Warsaw, Poland.
Centre of New Technologies, University of Warsaw, Banacha 2c, 02-097, Warsaw, Poland.
Sci Rep. 2017 Aug 9;7(1):7644. doi: 10.1038/s41598-017-08032-8.
Short modified oligonucleotides targeted at bacterial DNA or RNA could serve as antibacterial agents provided that they are efficiently taken up by bacterial cells. However, the uptake of such oligonucleotides is hindered by the bacterial cell wall. To overcome this problem, oligomers have been attached to cell-penetrating peptides, but the efficiency of delivery remains poor. Thus, we have investigated the ability of vitamin B to transport peptide nucleic acid (PNA) oligomers into cells of Escherichia coli and Salmonella Typhimurium. Vitamin B was covalently linked to a PNA oligomer targeted at the mRNA of a reporter gene expressing Red Fluorescent Protein. Cu-catalyzed 1,3-dipolar cycloaddition was employed for the synthesis of PNA-vitamin B conjugates; namely the vitamin B azide was reacted with PNA possessing the terminal alkyne group. Different types of linkers and spacers between vitamin B and PNA were tested, including a disulfide bond. We found that vitamin B transports antisense PNA into E. coli cells more efficiently than the most widely used cell-penetrating peptide (KFF)K. We also determined that the structure of the linker impacts the antisense effect. The results of this study provide the foundation for developing vitamin B as a carrier of PNA oligonucleotides into bacterial cells.
短的修饰寡核苷酸靶向细菌 DNA 或 RNA 可以作为抗菌剂，只要它们能有效地被细菌细胞吸收。然而，这些寡核苷酸的摄取受到细菌细胞壁的阻碍。为了克服这个问题，已经将低聚物与穿透细胞的肽连接起来，但是输送的效率仍然很差。因此，我们研究了维生素 B 将肽核酸（PNA）低聚物转运到大肠杆菌和鼠伤寒沙门氏菌细胞中的能力。维生素 B 与针对表达红色荧光蛋白的报告基因 mRNA 的 PNA 低聚物共价连接。Cu 催化的 1,3-偶极环加成反应用于合成 PNA-维生素 B 缀合物；即维生素 B 叠氮化物与具有末端炔基的 PNA 反应。测试了维生素 B 和 PNA 之间不同类型的连接子和间隔物，包括二硫键。我们发现维生素 B 将反义 PNA 转运到大肠杆菌细胞中的效率比最广泛使用的穿透细胞肽（KFF）K 更高。我们还确定了连接子的结构对反义效果的影响。这项研究的结果为开发维生素 B 作为 PNA 寡核苷酸进入细菌细胞的载体提供了基础。
