Intracellular uptake of modified oligonucleotide studied by two fluorescence techniques
Intracellular uptake of modified oligonucleotide studied by two fluorescence techniques
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DOI：
10.1002/bip.20055
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发表时间：
2004-05-01
期刊：
影响因子：
2.9
通讯作者：
Turpin, PY
中科院分区：
文献类型：
--
作者：
Kocisov치, E;Praus, P;Turpin, PY
相互作用，即，以阳离子多烯类抗生素-阿替霉素B 3-二甲氨基丙酰胺为载体，研究了合成的修饰反义寡核苷酸在B16黑色素瘤细胞中的细胞摄取和细胞内分布。用荧光四甲基罗丹明标记物标记含有等极性、非等排、基于膦酸酯的核苷酸间键3 '-O-P-CH 2-O-Y-的反义寡核苷酸-dT（15）寡聚体类似物。使用荧光显微成像技术观察寡核苷酸在细胞膜上的流动性及其在细胞内的分布。在最初的几个小时内，一个强烈的偏好染色的细胞核被found. Fluorescence寿命测量从细胞内环境（共焦激光显微荧光光谱仪，频域相位/调制技术在1至200 MHz的频率区域）产生两个光谱分量的4.9和1.4 ns的寿命，分别。虽然前一个成分与之前表征的荧光团与膜和/或细胞质中生物分子靶点结合的效应相关，但后一个成分是新观察到的，并讨论了其可能的起源。(C)2004 Wiley Periodicals，Inc.
Interaction, i.e., cellilar uptake and intracellular distribution, of synthetic modified antisense oligonucleotide with the B16 melanoma cell line was studied using cationic polyene antibiotic, amphotericin B 3-dimethylaminopropyl amide, as a carrier vector. The antisense oligo-nucleotide-dT(15) oligomer analogue containing isopolar, nonisosteric,phosphonate-based inter-nucleotide linkages 3'-O-P-CH2-O-Y-was labeled with,fluorescent tetramethylrhodamine marker. The oligonucleotide itinerancy across the cell membrane and its distribution inside the cell was visualized using fluorescence microimaging. During the first several hours a strong preference staining of the cell nucleus was found. Fluorescence lifetime measurements from the intracellular environment (confocal laser microspectrofluorimeter, frequency domain phase/modulation technique in 1 to 200 MHz frequency region) yielded two spectral components of 4.9 and 1.4 ns lifetime, respectively. While the former component correlates with the previously characterized effect of the fluorophore binding to biomolecular targets in membranes and/or cytoplasm, the latter component is newly observed and its possible origin is discussed. (C) 2004 Wiley Periodicals, Inc.