2025/07/19 by Baptiste Alberti, Séverine Vincent, Isabelle Stévant +6 · 1 voice · 1 citation
Biochemistry, Genetics and Molecular Biology · Medicine · #14-3-3 protein interactions #Biology #Cancer Cells and Metastasis #Cell #Cell Image Analysis Techniques #Computational biology #Enhancer #Evolutionary biology #Gene #Gene expression #Genetics #Multicellular organism
paper · doi:10.1093/nar/gkaf684
published in Nucleic Acids Research 53(14) (Oxford University Press)
openalex publication_date 2025/07/19 · openalex created_date 2025/10/10 · openalex updated_date 2026/08/05
Enhancers play an essential role in developmental processes by regulating the spatiotemporal expression of genes. Characterizing their spatiotemporal activity remains however an important challenge. Here we introduce a novel in vivo/in silico method for spatial single-cell enhancer-reporter assays (spatial-scERA) designed to reconstruct the spatial activity of candidate enhancer regions in parallel in multicellular organisms. Spatial-scERA integrates parallel reporter assays with single-cell RNA sequencing and spatial reconstruction using optimal transport, to map cell-type-specific enhancer activity at the single-cell level on a 3D virtual sample. We evaluated spatial-scERA in Drosophila embryos using 25 candidate enhancers, and validated the robustness of our reconstructions by comparing them to in situ hybridization. Remarkably, spatial-scERA faithfully reconstructed the spatial activity of these enhancers, even when the reporter construct was expressed in as few as 10 cells. Our results demonstrate the importance of integrating transcriptomic and spatial data for accurately predicting enhancer activity patterns in complex multicellular samples and linking enhancers to their potential target genes. Overall, spatial-scERA provides a scalable approach to map the spatial activity of enhancers at single-cell resolution without the need for imaging or a priori knowledge of embryology and can be applied to any multicellular organism amenable to transgenesis.