Hydrogel “Affinity Trap” for microRNAs with a Self-Assembling Fluorescent “Split-Probe” to Monitor Their Expression and Degradation

IF 5.5 2区化学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biomacromolecules Pub Date : 2025-07-14 DOI:10.1021/acs.biomac.5c00594

Sameen Yousaf , Bahareh Amirloo , Harmesh S. Aojula , David J. Clarke , Alberto Saiani , Andrew Irwin , Elena V. Bichenkova

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Abstract

The hydrogel “affinity trap” engineered here from peptides and nucleic acids into dynamic supramolecular structures offered the opportunity to measure physiological concentrations of tissue-specific microRNA expression and degradation, which are symptomatic for diseased cells and tissues. Hydrogel size-discriminating properties allowed to segregate microRNAs from complex biological media into a hydrogel matrix and entrap the target sequence via hybridization with a hydrogel-immobilized “capture” probe, where it could be detected through fluorescence quenching. We demonstrated the size-selective permeability of the hydrogel that provided a protective microenvironment for microRNAs and detection probes from cellular biological interference and afforded selective self-assembly and detection of oncogenic microRNA-21 (miR-21) in the presence of cell extracts, which is otherwise detrimental for detection in a gel-free solution. We were also able to monitor the degradation of unlabeled miR-21 by natural (RNase A and H) and synthetic (miR-21-RNase) ribonucleases and their synergistic actions, which could be potentially useful in the therapeutic knockdown of pathogenic RNAs.

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用自组装荧光“分裂探针”监测microrna表达和降解的水凝胶“亲和陷阱”。

从多肽和核酸到动态超分子结构的水凝胶“亲和陷阱”提供了测量组织特异性microRNA表达和降解的生理浓度的机会，这是患病细胞和组织的症状。水凝胶的大小区分特性允许将microrna从复杂的生物介质中分离到水凝胶基质中，并通过与水凝胶固定的“捕获”探针杂交捕获目标序列，在那里它可以通过荧光猝灭检测到。我们证明了水凝胶的大小选择性渗透性，它为microrna和检测探针提供了一个免受细胞生物干扰的保护微环境，并在细胞提取物存在的情况下选择性地自组装和检测致癌microRNA-21 (miR-21)，否则在无凝胶溶液中检测是有害的。我们还能够监测天然（RNase A和H）和合成（miR-21-RNase）核糖核酸酶对未标记的miR-21的降解及其协同作用，这可能在治疗性敲低致病性rna方面有潜在的用途。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biomacromolecules 化学-高分子科学

CiteScore

10.60

自引率

4.80%

发文量

417

审稿时长

1.6 months

期刊介绍： Biomacromolecules is a leading forum for the dissemination of cutting-edge research at the interface of polymer science and biology. Submissions to Biomacromolecules should contain strong elements of innovation in terms of macromolecular design, synthesis and characterization, or in the application of polymer materials to biology and medicine. Topics covered by Biomacromolecules include, but are not exclusively limited to: sustainable polymers, polymers based on natural and renewable resources, degradable polymers, polymer conjugates, polymeric drugs, polymers in biocatalysis, biomacromolecular assembly, biomimetic polymers, polymer-biomineral hybrids, biomimetic-polymer processing, polymer recycling, bioactive polymer surfaces, original polymer design for biomedical applications such as immunotherapy, drug delivery, gene delivery, antimicrobial applications, diagnostic imaging and biosensing, polymers in tissue engineering and regenerative medicine, polymeric scaffolds and hydrogels for cell culture and delivery.