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An aptamer-based, fluorescent and radionuclide dual-modality probe.
Biochimie ( IF 3.3 ) Pub Date : 2020-02-17 , DOI: 10.1016/j.biochi.2020.02.007
Gui-Xiong Zhang 1 , Yan-Lan Liu 2 , Min Yang 1 , Wen-Shan Huang 1 , Jie-Hua Xu 1
Affiliation  

Aptamers which are promising and effective molecular probes, can deliver either fluorescent materials or radionuclides to tumors. This study aimed to develop a novel both fluorescent and radionuclide dual-modality probe based on a truncated aptamer and evaluate its stability and binding affinities in vitro. The aptamer JHIT2 with binding specifically to HepG2 cells was previously generated by Cell-SELEX. Using mfold and RNAstructure software to predict the secondary structure folded by a middle random sequence to truncate the primer sequences at both ends of the aptamer JHIT2 to yield the aptamer JHIT2e, with a similar secondary structure to JHIT2 and the same specificity and affinity as JHIT2. Attaching carboxyfluorescein (FAM) readily to the aptamer JHIT2e and then attaching iodine-131 to the FAM moiety which has multiple sites for iodine labeling to develop a novel both fluorescent and radionuclide dual-modality probe, termed 131I-FAM-JHIT2e. Cell uptake and fluorescence imaging assays in vitro confirmed that 131I-FAM-JHIT2e had both FAM fluorescence signal and radio-activity signal and maintained specific binding ability to the human hepatoma cell line HepG2. This work formed a basis for aptamer-based, dual-modality imaging probe that contains both fluorescent and radionuclide tags, which also is potential for theranostics.

中文翻译:

基于适体的荧光和放射性核素双峰探针。

适体是有前途和有效的分子探针,可以将荧光物质或放射性核素递送至肿瘤。这项研究旨在开发一种新型的基于截短适体的荧光和放射性核素双峰探针,并在体外评估其稳定性和结合亲和力。先前通过Cell-SELEX生成了与HepG2细胞特异性结合的适体JHIT2。使用mfold和RNAstructure软件预测被中间随机序列折叠的二级结构,以截断适体JHIT2两端的引物序列以产生适体JHIT2e,其二级结构与JHIT2类似,并且特异性和亲和性与JHIT2相同。将羧基荧光素(FAM)轻松连接至适体JHIT2e,然后将碘131连接至具有多个碘标记位点的FAM部分,从而开发出一种新颖的荧光和放射性核素双峰探针,称为131I-FAM-JHIT2e。体外细胞摄取和荧光成像分析证实131I-FAM-JHIT2e同时具有FAM荧光信号和放射性信号,并保持了与人肝癌细胞系HepG2的特异性结合能力。这项工作为基于适体的双模态成像探针奠定了基础,该探针同时包含荧光和放射性核素标签,这对于治疗诊断学也具有潜在意义。体外细胞摄取和荧光成像分析证实131I-FAM-JHIT2e同时具有FAM荧光信号和放射性信号,并保持了与人肝癌细胞系HepG2的特异性结合能力。这项工作为基于适体的双模态成像探针奠定了基础,该探针同时包含荧光和放射性核素标签,这对于治疗诊断学也具有潜在意义。体外细胞摄取和荧光成像分析证实131I-FAM-JHIT2e同时具有FAM荧光信号和放射性信号,并保持了与人肝癌细胞系HepG2的特异性结合能力。这项工作为基于适体的双模态成像探针奠定了基础,该探针同时包含荧光和放射性核素标签,这对于治疗诊断学也具有潜在意义。
更新日期:2020-02-20
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