Department of Chemistry, College of Chemistry and Life Science, Beijing University of Technology, Beijing, China.
College of Biomass Science and Engineering, Healthy Food Evaluation Research Center, Sichuan University, Chengdu, China.
Methods Mol Biol. 2024;2822:65-75. doi: 10.1007/978-1-0716-3918-4_6.
We present a powerful method for direct mRNA detection based on ligation-based recognition and in situ amplification, capable of single-cell imaging mRNA at single-nucleotide and single-molecule resolution. Attributed to the use of Splint R ligase that can ligate padlock probe with RNA as target template, this method can efficiently detect mRNA in the absence of reverse transcription. This method enables spatial localization and correlation analysis of gene expression in single cells, which helps us to elucidate gene function and regulatory mechanisms.
我们提出了一种基于连接识别和原位扩增的强大的直接 mRNA 检测方法，能够以单核苷酸和单分子分辨率对单细胞内的 mRNA 进行成像。由于使用了 Splint R 连接酶，该酶可以将发夹探针与 RNA 作为靶模板连接，因此该方法可以在没有逆转录的情况下有效地检测 mRNA。该方法能够对单细胞中的基因表达进行空间定位和相关性分析，这有助于我们阐明基因功能和调控机制。
