当前位置: X-MOL 学术Anal. Chem. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Synthetic Host-Guest Assembly in Cells and Tissues: Fast, Stable, and Selective Bioorthogonal Imaging via Molecular Recognition.
Analytical Chemistry ( IF 6.7 ) Pub Date : 2018-08-27 00:00:00 , DOI: 10.1021/acs.analchem.8b01851
Ranjan Sasmal 1 , Nilanjana Das Saha 1, 2 , Meenakshi Pahwa 2 , Sushma Rao 3 , Divyesh Joshi 4 , Maneesha S Inamdar 4 , Vasu Sheeba 3 , Sarit S Agasti 1, 2, 5
Affiliation  

Bioorthogonal strategies are continuing to pave the way for new analytical tools in biology. Although a significant amount of progress has been made in developing covalent reaction based bioorthogonal strategies, balanced reactivity, and stability are often difficult to achieve from these systems. Alternatively, despite being kinetically beneficial, the development of noncovalent approaches that utilize fully synthetic and stable components remains challenging due to the lack of selectivity in conventional noncovalent interactions in the living cellular environment. Herein, we introduce a bioorthogonal assembly strategy based on a synthetic host–guest system featuring Cucurbit[7]uril (CB[7]) and adamantylamine (ADA). We demonstrate that highly selective and ultrastable host–guest interaction between CB[7] and ADA provides a noncovalent mechanism for assembling labeling agents, such as fluorophores and DNA, in cells and tissues for bioorthogonal imaging of molecular targets. Additionally, by combining with covalent reaction, we show that this CB[7]–ADA based noncovalent interaction enables simultaneous bioorthogonal labeling and multiplexed imaging in cells as well as tissue sections. Finally, we show that interaction between CB[7] and ADA fulfills the demands of specificity and stability that is required for assembling molecules in the complexities of a living cell. We demonstrate this by sensitive detection of metastatic cancer-associated cell surface protein marker as well as by showing the distribution and dynamics of F-actin in living cells.

中文翻译:


细胞和组织中的合成主客体组装:通过分子识别进行快速、稳定和选择性的生物正交成像。



生物正交策略正在继续为生物学中的新分析工具铺平道路。尽管在开发基于共价反应的生物正交策略方面已经取得了显着的进展,但这些系统通常难以实现平衡的反应性和稳定性。另外,尽管在动力学上是有益的,但由于活细胞环境中传统非共价相互作用缺乏选择性,利用完全合成和稳定成分的非共价方法的开发仍然具有挑战性。在此,我们介绍了一种基于以葫芦[7]脲(CB[7])和金刚胺(ADA)为特征的合成主客体系统的生物正交组装策略。我们证明,CB[7] 和 ADA 之间的高度选择性和超稳定的主客体相互作用提供了一种非共价机制,用于在细胞和组织中组装标记剂(例如荧光团和 DNA),以实现分子靶标的生物正交成像。此外,通过与共价反应相结合,我们表明这种基于 CB[7]-ADA 的非共价相互作用能够在细胞和组织切片中同时进行生物正交标记和多重成像。最后,我们证明 CB[7] 和 ADA 之间的相互作用满足了在活细胞的复杂性中组装分子所需的特异性和稳定性的要求。我们通过灵敏检测转移性癌症相关细胞表面蛋白标记物以及显示活细胞中 F-肌动蛋白的分布和动态来证明这一点。
更新日期:2018-08-27
down
wechat
bug