We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
ARTseq-FISH reveals position-dependent differences in gene expression of micropatterned mESCs.
- Authors
Hu, Xinyu; van Sluijs, Bob; García-Blay, Óscar; Stepanov, Yury; Rietrae, Koen; Huck, Wilhelm T. S.; Hansen, Maike M. K.
- Abstract
Differences in gene-expression profiles between individual cells can give rise to distinct cell fate decisions. Yet how localisation on a micropattern impacts initial changes in mRNA, protein, and phosphoprotein abundance remains unclear. To identify the effect of cellular position on gene expression, we developed a scalable antibody and mRNA targeting sequential fluorescence in situ hybridisation (ARTseq-FISH) method capable of simultaneously profiling mRNAs, proteins, and phosphoproteins in single cells. We studied 67 (phospho-)protein and mRNA targets in individual mouse embryonic stem cells (mESCs) cultured on circular micropatterns. ARTseq-FISH reveals relative changes in both abundance and localisation of mRNAs and (phospho-)proteins during the first 48 hours of exit from pluripotency. We confirm these changes by conventional immunofluorescence and time-lapse microscopy. Chemical labelling, immunofluorescence, and single-cell time-lapse microscopy further show that cells closer to the edge of the micropattern exhibit increased proliferation compared to cells at the centre. Together these data suggest that while gene expression is still highly heterogeneous position-dependent differences in mRNA and protein levels emerge as early as 12 hours after LIF withdrawal. Hu et al. describe ARTseq-FISH, a spatial omics method that enables multiplexed detection of mRNAs, proteins and phospho-proteins simultaneously and at the same resolution. The authors investigate gene expression changes in a heterogenous colony of micropatterned mESCs.
- Subjects
GENE expression; EMBRYONIC stem cells; CHEMICAL labeling; PHOSPHOPROTEINS
- Publication
Nature Communications, 2024, Vol 15, Issue 1, p1
- ISSN
2041-1723
- Publication type
Article
- DOI
10.1038/s41467-024-48107-5