核糖核酸
生物
原位杂交
寡核苷酸
DNA
分子生物学
基因表达
原位
信使核糖核酸
计算生物学
杂交探针
生物标志物
滚动圆复制
分子信标
基因
生物化学
聚合酶
化学
有机化学
作者
Meng Jiang,Kaipeng Wei,Meiqing Li,Chen Lin,Rongqin Ke
出处
期刊:RNA
日期:2023-02-22
卷期号:29 (6): 836-846
标识
DOI:10.1261/rna.079482.122
摘要
Although RNA plays a vital role in gene expression, it is less used as an in situ biomarker for clinical diagnostics than DNA and protein. This is mainly due to technical challenges caused by the low expression level and easy degradation of RNA molecules. To tackle this issue, methods that are sensitive and specific are needed. Here, we present an RNA single-molecule chromogenic in situ hybridization assay based on DNA probe proximity ligation and rolling circle amplification. When the DNA probes hybridize into close proximity to the RNA molecules, they form a V-shape structure and mediate the circularization of circle probes. Thus, our method was termed vsmCISH. We successfully applied our method to assess HER2 mRNA expression status in invasive breast cancer tissue and investigated the utility of albumin mRNA ISH for differentiating primary from metastatic liver cancer. The promising results on clinical samples indicate that our method has great potential for application in diagnosing diseases using RNA biomarkers.
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