Parallel analysis of tri-molecular biosynthesis with cell identity and function in single cells
Samuel C Kimmey1,2, Luciene Borges2, Reema Baskar2,3
1Department of Developmental Biology, Stanford University School of Medicine, Stanford, CA, 94305, USA.
Nature Communications
|March 14, 2019
Summary
We developed Simultaneous Overview of tri-Molecule Biosynthesis (SOM3B) to measure DNA, RNA, and protein synthesis in single cells. This method reveals how these processes coordinate during cell activation and differentiation.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Cellular identity and function are governed by DNA, RNA, and protein synthesis.
- Coordination of these biosynthesis pathways during transient cellular processes like activation and differentiation remains poorly understood.
Purpose of the Study:
- To introduce Simultaneous Overview of tri-Molecule Biosynthesis (SOM3B), a novel strategy for quantifying DNA, RNA, and protein synthesis in individual cells.
- To investigate the dynamics of de novo DNA, RNA, and protein synthesis in various human cell types and states.
Main Methods:
- Developed SOM3B, a molecular labeling and simultaneous detection strategy.
- Integrated SOM3B with antibody-based quantification for comprehensive analysis of transient cell states.
- Applied the method to transformed human cell lines, primary activated immune cells, and the human hematopoietic continuum.
Main Results:
- Enabled single-cell resolution quantification of simultaneous DNA, RNA, and protein synthesis.
- Revealed differential biosynthesis dynamics across various cellular contexts.
- Provided insights into molecular coordination during cell cycle progression and differentiation.
Conclusions:
- SOM3B offers a powerful approach to study coordinated molecular biosynthesis in single cells.
- The findings advance our understanding of cellular regulation during critical biological processes.
- This technique is applicable to diverse research areas in cell biology and immunology.
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