School of Chemistry and Food Engineering, Changsha University of Science and Technology, Changsha, 410114, China.
State Key Laboratory for Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, and Collaborative Research Center of Molecular Engineering for Theranostics, Hunan University, Changsha, 410082, China.
Angew Chem Int Ed Engl. 2019 Aug 19;58(34):11574-11585. doi: 10.1002/anie.201812449. Epub 2019 May 14.
Owing to its important physiological functions, especially as molecular biomarkers of diseases, RNA is an important focus of biomedicine and biochemical sensing. Signal amplification detection has been put forward because of the need for accurate identification of RNA at low expression levels, which is significant for the early diagnosis and therapy of malignant diseases. However, conventional amplification methods for RNA analysis depend on the use of enzymes, fixation of cells, and thermal cycling, which confine their performance to cell lysates or dead cells, thus the imaging of RNA in living cells remained until recently little explored. In recent years, the advance of isothermal amplification of nucleic acids has opened paths for meeting this need in living cells. This minireview tracks the development of in situ amplification assays for RNAs in living cells, and highlights the potential challenges facing this field, aiming to improve the development of in vivo isothermal amplification as well as usher in new frontiers in this fertile research area.
由于其重要的生理功能，尤其是作为疾病的分子生物标志物，RNA 是生物医学和生化传感的一个重要关注点。由于需要准确识别低表达水平的 RNA，因此提出了信号放大检测，这对于恶性疾病的早期诊断和治疗具有重要意义。然而，传统的 RNA 分析扩增方法依赖于酶的使用、细胞固定和热循环，这将其性能局限于细胞裂解物或死细胞，因此，直到最近，活细胞中 RNA 的成像仍未得到充分探索。近年来，核酸等温扩增的进展为满足这一需求开辟了道路。本综述跟踪了在活细胞中进行 RNA 原位扩增分析的发展，并强调了该领域面临的潜在挑战，旨在促进活体等温扩增的发展，并为这一富有成果的研究领域开辟新的前沿。
