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. 2024:2805:137-151.
doi: 10.1007/978-1-0716-3854-5_9.

Multi-labeling Live Imaging to Quantify Gene Expression Dynamics During Drosophila Embryonic Development

Sahla Syed  1 , Bomyi Lim  2
Affiliations

Multi-labeling Live Imaging to Quantify Gene Expression Dynamics During Drosophila Embryonic Development

Sahla Syed et al. Methods Mol Biol. 2024.

Abstract

Transcription in developing metazoans is inherently stochastic, involving transient and dynamic interactions among transcriptional machinery. A fundamental challenge with traditional techniques, including fixed-tissue protein and RNA staining, is the lack of temporal resolution. Quantifying kinetic changes in transcription can elucidate underlying mechanisms of interaction among regulatory modules. In this protocol, we describe the successful implementation of a combination of MS2/MCP and PP7/PCP systems in living Drosophila embryos to further our understanding of transcriptional dynamics during development. Our technique can be extended to visualize transcriptional activities of multiple genes or alleles simultaneously, characterize allele-specific expression of a target gene, and quantitatively analyze RNA polymerase II activity in a single-cell resolution.

Keywords: Development; Drosophila embryos; MS2; PP7; Quantitative live imaging; Transcription.

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References

    1. Coppola CJ, Ramaker RC, Mendenhall EM (2016) Identification and function of enhancers in the human genome. Hum Mol Genet 25:R190–R197 - DOI - PubMed
    1. Levine M (2010) Transcriptional enhancers in animal development and evolution. Curr Biol 20:R754–R763 - DOI - PubMed - PMC
    1. Coulon A, Chow CC, Singer RH, Larson DR (2013) Eukaryotic transcriptional dynamics: from single molecules to cell populations. Nat Rev Genet 14:572–584. https://doi.org/10.1038/NRG3484 - DOI - PubMed
    1. Hocine S, Raymond P, Zenklusen D et al (2013) Single-molecule analysis of gene expression using two-color RNA labeling in live yeast. Nat Methods 10:119–121. https://doi.org/10.1038/nmeth.2305 - DOI - PubMed
    1. Larson DR, Zenklusen D, Wu B et al (2011) Real-time observation of transcription initiation and elongation on an endogenous yeast gene. Science 332:475. https://doi.org/10.1126/SCIENCE.1202142 - DOI - PubMed - PMC

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