Single embryo-resolution quantitative analysis of reporters permits multiplex spatial cis-regulatory analysis

Dev Biol. 2017 Feb 15;422(2):92-104. doi: 10.1016/j.ydbio.2017.01.010. Epub 2017 Jan 16.

Abstract

Cis-regulatory modules (CRMs) control spatiotemporal gene expression patterns in embryos. While measurement of quantitative CRM activities has become efficient, measurement of spatial CRM activities still relies on slow, one-by-one methods. To overcome this bottleneck, we have developed a high-throughput method that can simultaneously measure quantitative and spatial CRM activities. The new method builds profiles of quantitative CRM activities measured at single-embryo resolution in many mosaic embryos and uses these profiles as a 'fingerprint' of spatial patterns. We show in sea urchin embryos that the new method, Multiplex and Mosaic Observation of Spatial Information encoded in Cis-regulatory modules (MMOSAIC), can efficiently predict spatial activities of new CRMs and can detect spatial responses of CRMs to gene perturbations. We anticipate that MMOSAIC will facilitate systems-wide functional analyses of CRMs in embryos.

Keywords: Cis-regulatory; Embryo; Gene regulatory network; High-throughput; Mosaic; Reporter assay; Sea urchin; Spatial.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA / genetics
  • DNA Barcoding, Taxonomic / methods*
  • Embryo, Nonmammalian / embryology
  • Gene Expression Regulation, Developmental / genetics*
  • Genomics / methods*
  • High-Throughput Screening Assays / methods*
  • RNA / genetics
  • Regulatory Elements, Transcriptional / genetics*
  • Sea Urchins / embryology*
  • Sequence Analysis, DNA / methods
  • Transcription Factors / metabolism

Substances

  • Transcription Factors
  • RNA
  • DNA