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Dis Model Mech. 2017 Jul 1;10(7):923-929. doi: 10.1242/dmm.030163. Epub 2017 May 11.

Parallel imaging of Drosophila embryos for quantitative analysis of genetic perturbations of the Ras pathway.

Author information

1
Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA.
2
Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, NJ 08544, USA.
3
School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA.
4
Department of Statistics, Stanford University, Stanford, CA 94305, USA.
5
Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA stas@princeton.edu hang.lu@gatech.edu.
6
School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA stas@princeton.edu hang.lu@gatech.edu.

Abstract

The Ras pathway patterns the poles of the Drosophila embryo by downregulating the levels and activity of a DNA-binding transcriptional repressor Capicua (Cic). We demonstrate that the spatiotemporal pattern of Cic during this signaling event can be harnessed for functional studies of mutations in the Ras pathway in human diseases. Our approach relies on a new microfluidic device that enables parallel imaging of Cic dynamics in dozens of live embryos. We found that although the pattern of Cic in early embryos is complex, it can be accurately approximated by a product of one spatial profile and one time-dependent amplitude. Analysis of these functions of space and time alone reveals the differential effects of mutations within the Ras pathway. Given the highly conserved nature of Ras-dependent control of Cic, our approach provides new opportunities for functional analysis of multiple sequence variants from developmental abnormalities and cancers.

KEYWORDS:

Capicua; Microfluidics; Mutations; Ras; SVD

PMID:
28495673
PMCID:
PMC5536913
DOI:
10.1242/dmm.030163
[Indexed for MEDLINE]
Free PMC Article

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