دورية أكاديمية
Visualization of Individual RNA Molecules by Proximity Ligation-Based Chromogenic In Situ Hybridization Assay.
| العنوان: | Visualization of Individual RNA Molecules by Proximity Ligation-Based Chromogenic In Situ Hybridization Assay. |
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| المؤلفون: | Xia X; School of Medicine, Huaqiao University, Xiamen, Fujian, China., Jiang M; School of Medicine, Huaqiao University, Xiamen, Fujian, China.; College of Materials Science and Engineering, Huaqiao University, Xiamen, Fujian, China., Lin C; School of Medicine, Huaqiao University, Xiamen, Fujian, China., Ke R; School of Medicine, Huaqiao University, Xiamen, Fujian, China. rke@hqu.edu.cn. |
| المصدر: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2024; Vol. 2822, pp. 143-156. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Humana Press Country of Publication: United States NLM ID: 9214969 Publication Model: Print Cited Medium: Internet ISSN: 1940-6029 (Electronic) Linking ISSN: 10643745 NLM ISO Abbreviation: Methods Mol Biol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Totowa, NJ : Humana Press Original Publication: Clifton, N.J. : Humana Press, |
| مواضيع طبية MeSH: | In Situ Hybridization*/methods , RNA*/genetics , RNA*/analysis, Humans ; Chromogenic Compounds/chemistry ; Colorimetry/methods ; Single Molecule Imaging/methods |
| مستخلص: | RNA in situ hybridization reveals the abundance and location of gene expression in cells or tissues, providing a technical basis for the clinical diagnosis of diseases. In this chapter, we show a "V" shape probe-mediated single-molecule chromogenic in situ hybridization (vsmCISH) technique for bright-field visualization of individual RNA molecules. In our method, several pairs of target hybridization probes are hybridized to RNA molecules and each probe pair forms a "V" shape overhang. The overhang oligonucleotides then mediated the proximity ligation to form DNA circles, followed by rolling circle amplification for signal enhancement and enzyme-catalyzed chromogenic reaction-based readout. The colorimetric assay avoids problems such as photobleaching and autofluorescence of current fluorescent in situ hybridization-based single-molecule RNA detection techniques. Furthermore, the relatively straightforward protocol makes the method useful for biological research and clinical diagnosis applications. (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.) |
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| فهرسة مساهمة: | Keywords: Bright-field microscopy; Proximity ligation; RNA in situ hybridization; Rolling circle amplification |
| المشرفين على المادة: | 63231-63-0 (RNA) 0 (Chromogenic Compounds) |
| تواريخ الأحداث: | Date Created: 20240622 Date Completed: 20240622 Latest Revision: 20240622 |
| رمز التحديث: | 20240623 |
| DOI: | 10.1007/978-1-0716-3918-4_11 |
| PMID: | 38907917 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1940-6029 |
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| DOI: | 10.1007/978-1-0716-3918-4_11 |