Format

Send to

Choose Destination
Nat Commun. 2018 Jun 19;9(1):2384. doi: 10.1038/s41467-018-04761-0.

Immuno-detection by sequencing enables large-scale high-dimensional phenotyping in cells.

Author information

1
Department of Molecular Developmental Biology, Radboud Institute for Molecular Life Sciences, Radboud University, PO Box 9101, 6500 HB, Nijmegen, The Netherlands.
2
Department of Human Genetics, Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, PO Box 9101, 6500 HB, Nijmegen, The Netherlands.
3
The Oncode Institute, Department of Molecular Biology, Radboud Institute for Molecular Life Sciences, Radboud University, PO Box 9101, 6500 HB, Nijmegen, The Netherlands.
4
Biolegio BV, PO Box 91, 6500 AB, Nijmegen, The Netherlands.
5
Department of Molecular Developmental Biology, Radboud Institute for Molecular Life Sciences, Radboud University, PO Box 9101, 6500 HB, Nijmegen, The Netherlands. k.mulder@science.ru.nl.

Abstract

Cell-based small molecule screening is an effective strategy leading to new medicines. Scientists in the pharmaceutical industry as well as in academia have made tremendous progress in developing both large-scale and smaller-scale screening assays. However, an accessible and universal technology for measuring large numbers of molecular and cellular phenotypes in many samples in parallel is not available. Here we present the immuno-detection by sequencing (ID-seq) technology that combines antibody-based protein detection and DNA-sequencing via DNA-tagged antibodies. We use ID-seq to simultaneously measure 70 (phospho-)proteins in primary human epidermal stem cells to screen the effects of ~300 kinase inhibitor probes to characterise the role of 225 kinases. The results show an association between decreased mTOR signalling and increased differentiation and uncover 13 kinases potentially regulating epidermal renewal through distinct mechanisms. Taken together, our work establishes ID-seq as a flexible solution for large-scale high-dimensional phenotyping in fixed cell populations.

Supplemental Content

Full text links

Icon for Nature Publishing Group Icon for PubMed Central
Loading ...
Support Center