Single mRNA Molecule Detection in Drosophila

Shawn Little (1) , Thomas Gregor (2)

1 Perelman School of Medicine
2 Lewis-Sigler Institute for Integrative Genomics

Single molecule fluorescent in situ hybridization (smFISH) enables quantitative measurements of gene expression and mRNA localization. The technique is increasingly popular for analysis of cultured cells but is not widely applied to intact organisms. Here, we describe a method for labeling and detection of single mRNA molecules in whole embryos of the fruit fly Drosophila melanogaster. This method permits measurements of gene expression in absolute units, enabling new studies of transcriptional mechanisms underlying precision and reproducibility in cell specification.

Keywords: Single molecule; FISH; Quantitation; Fluorescence imaging; Drosophila

Domains: Biological Physics [physics.bio-ph]; Animal genetics

Shawn Little, Thomas Gregor. Single mRNA Molecule Detection in Drosophila. RNA Detection, 1649, Springer New York, pp.127-142, 2018, Methods in Molecular Biology, ⟨10.1007/978-1-4939-7213-5_8⟩. ⟨pasteur-03869357⟩