High-throughput protocol for studying pri-miRNA processing using randomized sequences
Summary: The Microprocessor complex is crucial in microRNA (miRNA) biogenesis, as it processes primary miRNAs (pri-miRNAs) into precursor miRNAs. Here, we present a high-throughput, radioisotope-free protocol for studying pri-miRNA processing using randomized sequences. We describe steps for randomi...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2024-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723007499 |
| _version_ | 1797388715510726656 |
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| author | Thi Nhu-Y Le Tuan Anh Nguyen |
| author_facet | Thi Nhu-Y Le Tuan Anh Nguyen |
| author_sort | Thi Nhu-Y Le |
| collection | DOAJ |
| description | Summary: The Microprocessor complex is crucial in microRNA (miRNA) biogenesis, as it processes primary miRNAs (pri-miRNAs) into precursor miRNAs. Here, we present a high-throughput, radioisotope-free protocol for studying pri-miRNA processing using randomized sequences. We describe steps for randomized substrate preparation, protein purification, processing assays, and DNA library construction for sequencing. This technique explores pri-miRNA processing, uncovers key RNA elements, and illuminates gene expression regulation. However, its efficacy may be constrained by data analysis complexity and the requirement for specialized equipment.For complete details on the use and execution of this protocol, please refer to Nguyen et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-08T22:44:49Z |
| format | Article |
| id | doaj.art-f1646fd1daa04265b360b93645b53e27 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-08T22:44:49Z |
| publishDate | 2024-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-f1646fd1daa04265b360b93645b53e272023-12-17T06:41:48ZengElsevierSTAR Protocols2666-16672024-03-0151102782High-throughput protocol for studying pri-miRNA processing using randomized sequencesThi Nhu-Y Le0Tuan Anh Nguyen1Division of Life Science, The Hong Kong University of Science & Technology, Hong Kong, ChinaDivision of Life Science, The Hong Kong University of Science & Technology, Hong Kong, China; Corresponding authorSummary: The Microprocessor complex is crucial in microRNA (miRNA) biogenesis, as it processes primary miRNAs (pri-miRNAs) into precursor miRNAs. Here, we present a high-throughput, radioisotope-free protocol for studying pri-miRNA processing using randomized sequences. We describe steps for randomized substrate preparation, protein purification, processing assays, and DNA library construction for sequencing. This technique explores pri-miRNA processing, uncovers key RNA elements, and illuminates gene expression regulation. However, its efficacy may be constrained by data analysis complexity and the requirement for specialized equipment.For complete details on the use and execution of this protocol, please refer to Nguyen et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723007499Sequence analysisGene Expression |
| spellingShingle | Thi Nhu-Y Le Tuan Anh Nguyen High-throughput protocol for studying pri-miRNA processing using randomized sequences STAR Protocols Sequence analysis Gene Expression |
| title | High-throughput protocol for studying pri-miRNA processing using randomized sequences |
| title_full | High-throughput protocol for studying pri-miRNA processing using randomized sequences |
| title_fullStr | High-throughput protocol for studying pri-miRNA processing using randomized sequences |
| title_full_unstemmed | High-throughput protocol for studying pri-miRNA processing using randomized sequences |
| title_short | High-throughput protocol for studying pri-miRNA processing using randomized sequences |
| title_sort | high throughput protocol for studying pri mirna processing using randomized sequences |
| topic | Sequence analysis Gene Expression |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723007499 |
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