Quantification of purified endogenous miRNAs with high sensitivity and specificity
| العنوان: | Quantification of purified endogenous miRNAs with high sensitivity and specificity |
|---|---|
| المؤلفون: | Heesoo Uhm, Ji-Joon Song, Cherlhyun Jeong, Jiyoung Goo, Soomin Son, V. Narry Kim, Yoonseok Jung, Sungchul Hohng, Soochul Shin, Minseok Song |
| المصدر: | Nature Communications Nature Communications, Vol 11, Iss 1, Pp 1-8 (2020) |
| بيانات النشر: | Nature Publishing Group UK, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | 0301 basic medicine, Science, General Physics and Astronomy, Endogeny, Computational biology, Biology, 010402 general chemistry, 01 natural sciences, General Biochemistry, Genetics and Molecular Biology, Article, Imaging, Cell Line, 03 medical and health sciences, microRNA, medicine, Humans, Differential expression, Thermus, Gene, Multidisciplinary, medicine.diagnostic_test, Base Sequence, General Chemistry, Argonaute, Thermus thermophilus, biology.organism_classification, 0104 chemical sciences, MicroRNAs, 030104 developmental biology, miRNAs, Imaging technique, Fluorescence in situ hybridization |
| الوصف: | MicroRNAs (miRNAs) are short (19–24 nt) non-coding RNAs that suppress the expression of protein coding genes at the post-transcriptional level. Differential expression profiles of miRNAs across a range of diseases have emerged as powerful biomarkers, making a reliable yet rapid profiling technique for miRNAs potentially essential in clinics. Here, we report an amplification-free multi-color single-molecule imaging technique that can profile purified endogenous miRNAs with high sensitivity, specificity, and reliability. Compared to previously reported techniques, our technique can discriminate single base mismatches and single-nucleotide 3′-tailing with low false positive rates regardless of their positions on miRNA. By preloading probes in Thermus thermophilus Argonaute (TtAgo), miRNAs detection speed is accelerated by more than 20 times. Finally, by utilizing the well-conserved linearity between single-molecule spot numbers and the target miRNA concentrations, the absolute average copy numbers of endogenous miRNA species in a single cell can be estimated. Thus our technique, Ago-FISH (Argonaute-based Fluorescence In Situ Hybridization), provides a reliable way to accurately profile various endogenous miRNAs on a single miRNA sensing chip. MicroRNAs are potentially powerful biomarkers, though clinical use requires rapid and reliable profiling. Here the authors report amplification-free multicolour single-molecule imaging with single base mismatch sensitivity. |
| اللغة: | English |
| تدمد: | 2041-1723 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::84369d3732f93eecee200b20424d4735 http://europepmc.org/articles/PMC7699633 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....84369d3732f93eecee200b20424d4735 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 20411723 |
|---|