Narayanaswamy Nagarjun, Kumar Manoj, Das Sadhan, Sharma Rahul, Samanta Pralok K, Pati Swapan K, Dhar Suman K, Kundu Tapas K, Govindaraju T
Bioorganic Chemistry Laboratory, New Chemistry Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Jakkur P.O., Bangalore 560064, Karnataka, India.
Transcription and Disease Laboratory, Molecular Biology and Genetics Unit, Jawaharlal Nehru Centre for Advanced Scientific Research, Jakkur P.O., Bangalore 560064, Karnataka, India.
Sci Rep. 2014 Sep 25;4:6476. doi: 10.1038/srep06476.
Sequence-specific recognition of DNA by small turn-on fluorescence probes is a promising tool for bioimaging, bioanalytical and biomedical applications. Here, the authors report a novel cell-permeable and red fluorescent hemicyanine-based thiazole coumarin (TC) probe for DNA recognition, nuclear staining and cell cycle analysis. TC exhibited strong fluorescence enhancement in the presence of DNA containing AT-base pairs, but did not fluoresce with GC sequences, single-stranded DNA, RNA and proteins. The fluorescence staining of HeLa S3 and HEK 293 cells by TC followed by DNase and RNase digestion studies depicted the selective staining of DNA in the nucleus over the cytoplasmic region. Fluorescence-activated cell sorting (FACS) analysis by flow cytometry demonstrated the potential application of TC in cell cycle analysis in HEK 293 cells. Metaphase chromosome and malaria parasite DNA imaging studies further confirmed the in vivo diagnostic and therapeutic applications of probe TC. Probe TC may find multiple applications in fluorescence spectroscopy, diagnostics, bioimaging and molecular and cell biology.
通过小分子开启荧光探针实现对DNA的序列特异性识别,是一种在生物成像、生物分析和生物医学应用方面颇具前景的工具。在此,作者报道了一种新型的、可穿透细胞的、基于半花菁的红色荧光噻唑香豆素(TC)探针,用于DNA识别、细胞核染色和细胞周期分析。在含有AT碱基对的DNA存在时,TC表现出强烈的荧光增强,但与GC序列、单链DNA、RNA和蛋白质不发生荧光反应。用TC对HeLa S3和HEK 293细胞进行荧光染色,随后进行DNA酶和RNA酶消化研究,结果表明细胞核中的DNA相对于细胞质区域具有选择性染色。通过流式细胞术进行的荧光激活细胞分选(FACS)分析证明了TC在HEK 293细胞的细胞周期分析中的潜在应用。中期染色体和疟原虫DNA成像研究进一步证实了探针TC在体内诊断和治疗方面的应用。探针TC可能在荧光光谱、诊断、生物成像以及分子和细胞生物学中找到多种应用。