Journal article
Protocol to quantify the rate of RNA polymerase II elongation with MS2/MCP- and PP7/PCP-based live-imaging systems in early Drosophila embryos
STAR protocols, v 5(2), p103099
21 Jun 2024
PMID: 38824639
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
The MS2-PP7 two-color live-imaging system provides insights into the spatiotemporal dynamics of nascent transcripts at tagged loci. Here, we present a protocol to quantitatively measure the rate of RNA polymerase II elongation for each actively transcribing nucleus in living Drosophila embryos. The elongation rate is calculated by measuring the effective distance and the time elapsed between MS2 and PP7 trajectories. We describe steps for preparing embryo samples, performing live imaging, and measuring the elongation rate.
For complete details on the use and execution of this protocol, please refer to Keller et al.1
[Display omitted]
•A protocol for measuring the elongation rate of RNA Pol II in Drosophila embryos•Steps for using MS2 and PP7 live imaging to measure elongation rates in single cells•Steps to use “leading-edge” and “cross-correlation” methods to measure elongation rates
Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
The MS2-PP7 two-color live-imaging system provides insights into the spatiotemporal dynamics of nascent transcripts at tagged loci. Here, we present a protocol to quantitatively measure the rate of RNA polymerase II elongation for each actively transcribing nucleus in living Drosophila embryos. The elongation rate is calculated by measuring the effective distance and the time elapsed between MS2 and PP7 trajectories. We describe steps for preparing embryo samples, performing live imaging, and measuring the elongation rate.
Metrics
2 Record Views
Details
- Title
- Protocol to quantify the rate of RNA polymerase II elongation with MS2/MCP- and PP7/PCP-based live-imaging systems in early Drosophila embryos
- Creators
- Hao Deng - University of PennsylvaniaBomyi Lim - University of Pennsylvania
- Publication Details
- STAR protocols, v 5(2), p103099
- Publisher
- Elsevier Inc
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- School of Biomedical Engineering, Science, and Health Systems
- Web of Science ID
- WOS:001248570400002
- Scopus ID
- 2-s2.0-85194468399
- Other Identifier
- 991022138680704721
UN Sustainable Development Goals (SDGs)
This publication has contributed to the advancement of the following goals:
InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Web of Science research areas
- Biochemical Research Methods