合成生物学 ›› 2022, Vol. 3 ›› Issue (2): 369-384.DOI: 10.12211/2096-8280.2021-060
武伟红1, 李炜2, 张先恩3, 崔宗强2
收稿日期:
2021-05-13
修回日期:
2021-09-06
出版日期:
2022-04-30
发布日期:
2022-05-11
通讯作者:
崔宗强
作者简介:
基金资助:
Weihong WU1, Wei LI2, Xian’en ZHANG3, Zongqiang CUI2
Received:
2021-05-13
Revised:
2021-09-06
Online:
2022-04-30
Published:
2022-05-11
Contact:
Zongqiang CUI
摘要:
合成生物学的迅速发展为分子荧光标记与生物成像技术提供了新的机遇。基于合成生物学原理,可以建立材料生物合成新方法,开发性能优异的荧光纳米材料和探针,发展新的荧光成像技术。合成生物学应用于生物荧光成像,多涉及荧光材料与探针的设计合成、对生物靶标分子进行定点改造和修饰、荧光探针和靶标分子的可控时空耦合等以实现生物分子的精准特异性标记。这些荧光纳米材料和生物分子标记技术可应用于细胞内分子的荧光标记、成像和动态示踪,可视化解析相关的关键分子事件,从而深入揭示细胞内分子运动机制和病原致病机理等。本文主要综述了近年来合成生物学技术在生物荧光成像方面的应用,包括利用合成生物学技术合成量子点等荧光纳米材料与探针、对蛋白质和核酸分子的精准标记及其用于病毒荧光成像和示踪。最后,也对该领域面临的问题如荧光杂合生物材料可控合成、分子原位多重标记等进行了探讨和展望。合成生物学与荧光成像技术的交叉融合,将推动荧光成像技术发展和进步,并拓展合成生物学的研究领域。
中图分类号:
武伟红, 李炜, 张先恩, 崔宗强. 合成生物学与荧光成像技术[J]. 合成生物学, 2022, 3(2): 369-384.
Weihong WU, Wei LI, Xian’en ZHANG, Zongqiang CUI. Synthetic biology for fluorescent bioimaging[J]. Synthetic Biology Journal, 2022, 3(2): 369-384.
荧光材料 | 大小 | 荧光强度 | 荧光量子效率 | 荧光寿命 |
---|---|---|---|---|
量子点 | 2~20 nm | +++++ | 0.1~0.85 | 20 min |
荧光蛋白 | 4 nm | + | 0.79 | 100 ms |
荧光RNA | 40 bp | +++ | 0.30~0.70 | > 15 min |
表1 不同荧光材料性能参数[74-76]
Tab. 1 Properties of different fluorescent materials [74-76]
荧光材料 | 大小 | 荧光强度 | 荧光量子效率 | 荧光寿命 |
---|---|---|---|---|
量子点 | 2~20 nm | +++++ | 0.1~0.85 | 20 min |
荧光蛋白 | 4 nm | + | 0.79 | 100 ms |
荧光RNA | 40 bp | +++ | 0.30~0.70 | > 15 min |
图5 核酸适配子Corn荧光基团成像单细胞中聚合酶活性示意图[87](转入报告质粒的HEK293T细胞加入荧光团DFHO可产生黄色荧光,加入RNA聚合酶抑制剂放线菌素D后荧光消失)
Fig. 5 Schematic diagram for imaging polymerase activity in single cells through aptamer corn fluorophore[87](HEK293T cells transformed with the reporter plasmid could produce yellow fluorescence by adding fluorophore DFHO, but the fluorescence disappears after adding the RNA polymerase inhibitor actinomycin D.)
图7 荧光RNA复合体在HeLa细胞成像[76](合成染料HBC的一系列类似物与Pepper结合可发出不同颜色荧光,HBC类似物与Pepper结合物在HeLa细胞中进行成像,用空载细胞做对比,在共聚焦显微镜和双光子显微镜下进行成像观察)
Fig. 7 Schematic diagram for imaging of the fluorescent RNA complex in HeLa cells[76](A series of analogues of synthetic dye HBC combined with Pepper can emit fluorescence with different colors. HBC analogues were combined with Pepper in HeLa cells and are compared with mock cells. Imaging observation is carried out with confocal microscope and two-photon microscope.)
图9 流感病毒脱壳过程实时动态示踪[75][QD625在电镜下又黑又圆,而QD705为三角形。用QD625(红色)和QD705(绿色)分别标记vRNP,结果显示在脱壳过程中QD625和QD705分别释放。IAV释放的vRNPs经过三阶段的主动运输到达细胞核,在细胞核经历两种扩散模式]
Fig. 9 Real-time dynamic tracking of the influenza virus unpacking process[75][Under electron microscope, QD625 is black and round, while QD705 is triangular. vRNP is labeled with QD625 (red) and QD705 (green) respectively, and the results showed that QD625 and QD705 are released during the hulling process, respectively. IAV-released vRNPs released from IAV reach the nucleus through a three-stage active transport, where they undergo two diffusion modes.]
图11 SV40-QD VLPs用于靶向小鼠动脉粥样硬化斑块成像[100](SV40 VLPs 含有动脉粥样硬化靶向单元,凝血酶抑制剂并可以封装QD800。FH-QDs可靶向集中在小鼠体内动脉粥样硬化形成区)
Fig. 11 SV40-QD VLPs for imaging atherosclerotic plaques in targeted mice[100](SV40 VLPs contain atherosclerotic targeting units, thrombin inhibitors, which can encapsulate as QD800. FH-QDS can target atherosclerotic areas in mice.)
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