Lin Chen, Xiao Zhehao, Zhang Xinya, Wu Guangxiong
School of Medicine and School of Biomedical Sciences, Huaqiao University, Quanzhou, 362021, Fujian, China.
School of Medicine and School of Biomedical Sciences, Huaqiao University, Quanzhou, 362021, Fujian, China.
Biochem Biophys Res Commun. 2022 Jun 25;610:30-34. doi: 10.1016/j.bbrc.2022.04.034. Epub 2022 Apr 10.
Circular RNAs (circRNAs) are a class of noncoding RNAs generated by a specific type of RNA alternative splicing called backsplicing through various mechanisms. Recently, thousands of circRNAs have been identified by high-throughput RNA sequencing technologies and bioinformatics analysis. However, the functions of the majority have not been fully elucidated yet. Different tools, such as in situ hybridization, can help visualize the spatial temporal distribution of circRNA molecules, thus assisting the understanding of their biological and physiological functions. Here, we present a simple and straightforward method based on padlock probe hybridization and rolling circle amplification (RCA) for in situ detection of circRNAs. We compared our method with the commercially available BaseScope assay for the detection of Cdr1as in the mouse brain tissue. The result showed that the two methods have achieved comparable detection efficiency, thus demonstrating our padlock probe assay as an alternative yet simple circRNA in situ detection method for the research community.
环状RNA（circRNAs）是一类非编码RNA，通过各种机制由一种名为反向剪接的特定类型的RNA可变剪接产生。最近，通过高通量RNA测序技术和生物信息学分析已鉴定出数千种circRNAs。然而，大多数circRNAs的功能尚未完全阐明。不同的工具，如原位杂交，有助于可视化circRNA分子的时空分布，从而有助于理解它们的生物学和生理功能。在此，我们提出了一种基于锁式探针杂交和滚环扩增（RCA）的简单直接的方法，用于原位检测circRNAs。我们将我们 的方法与市售的BaseScope检测法在小鼠脑组织中检测Cdr1as进行了比较。结果表明，这两种方法具有相当的检测效率，从而证明我们的锁式探针检测法是一种可供研究界使用的、简单的circRNA原位检测替代方法。
