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Protocol for quantitative analysis of RNA 3'-end processing induced by disassociated subunits using
Jie Huang1, Lijun Bao1, Junyi Zhu1
1Key Laboratory of Cell Proliferation and Differentiation of the Ministry of Education, School of Life Sciences, Peking-Tsinghua Center for Life Sciences, Peking University, Beijing 100871, China.
STAR Protocols
|June 17, 2023
Summary
This study introduces a new method for analyzing chromatin-associated RNA sequencing (RNA-seq) data to identify readthrough transcripts. The protocol helps researchers understand RNA processing and gene expression in detail.
Area of Science:
- Molecular Biology
- Genomics
- Transcriptomics
Background:
- Chromatin-associated RNA sequencing (RNA-seq) allows for the study of RNA processing.
- Identifying readthrough transcripts is crucial for understanding gene regulation.
Purpose of the Study:
- To present an experimental strategy and computational pipeline for processing chromatin-associated RNA-seq data.
- To detect and quantify readthrough transcripts.
Main Methods:
- Construction of degron mouse embryonic stem cells.
- Development of a computational pipeline for RNA-seq data analysis.
- Detection and quantification of readthrough genes.
Main Results:
- The study provides a detailed protocol for analyzing chromatin-associated RNA-seq data.
- The method enables the detection and quantification of readthrough transcripts.
Conclusions:
- The presented protocol is adaptable for various biological contexts and nascent RNA-seq techniques.
- This approach enhances the characterization of RNA processing and gene expression.
