单个RNA分子的染色体可视化In Situ通过滚动圆放大与性酸酶进行双重能力
Ruolan Huang1, Kun Liu2, Chensirong Yue1
1Engineering Research Centre of Molecular Medicine of Ministry of Education, Key Laboratory of Fujian Molecular Medicine, Key Laboratory of Precision Medicine and Molecular Diagnosis of Fujian Universities, School of Medicine, Huaqiao University, Quanzhou, Fujian 362021, China.
Analytical chemistry
|July 31, 2023
概括
我们开发了一种用户友好的方法,用于在单分子水平上使用性酸酶 (ALP) 和滚动圆放大来可视化mRNA. 这种技术提供了敏感和特定的RNA检测,克服了背景问题,改善了诊断.
科学领域:
- 分子生物学分子生物学
- 生物技术是生物技术.
- 显微镜的使用方法
背景情况:
- 在单分子分辨率下可视化空间基因表达是成像中的一个重大挑战.
- 现有的RNA检测方法可能是复杂的或遭受背景干扰.
研究的目的:
- 利用光学显微镜开发一种用户友好,敏感和特定的单分子RNA可视化方法.
- 为了实现双重RNA检测,同时分析多个基因标.
主要方法:
- 开发了一种新的方法,利用滚动圆放大与酸酶 (SA-ALP) 进行RNA检测.
- 采用快红色染色用于信号生成和图像分析.
- 调整了系统用于双重测定,使用性酸酶与胡卜氧化酶进行双色检测.
主要成果:
- 实现了高度选择性和敏感的RNA检测,与基于光的测试和RNAscope可比.
- 即使在富含黑色素的组织中,也成功量化了RNA斑点.
- 成功执行双重测试,在单细胞中同时可视化和量化两个不同的mRNA目标.
结论:
- 开发的SA-ALP方法为单分子RNA可视化提供了一种多功能,用户友好和敏感的方法.
- 这种技术克服了背景干扰,适用于具有挑战性的样品,如富含黑色素的组织.
- 该方法的低硬件要求和对双重测试的适应性可以使资源有限的机构能够进行RNA研究和诊断.
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