Decoding Expression Dynamics of Protein and Transcriptome at the Single-Cell Level in Paired Picoliter Chambers
Xing Xu1, Mingxia Zhang1,2, Xuebing Zhang2
1Collaborative Innovation Center of Chemistry for Energy Materials, The MOE Key Laboratory of Spectrochemical Analysis and Instrumentation, State Key Laboratory of Physical Chemistry of Solid Surfaces, Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, Fujian 361005, China.
We developed multi-Paired-seq, a cost-effective platform for simultaneous single-cell mRNA and protein analysis. This method enhances cell utilization and accuracy, providing new insights into cellular heterogeneity and gene-protein correlations.
Area of Science:
- Molecular Biology
- Genomics
- Proteomics
Background:
- Simultaneous single-cell analysis of mRNA and protein is crucial for understanding cellular heterogeneity and gene-protein dynamics.
- Existing methods face challenges in cell utilization, accuracy, and cost-effectiveness.
Purpose of the Study:
- To present multi-Paired-seq, a novel platform for high-throughput, cost-effective, simultaneous analysis of mRNAs and membrane proteins at the single-cell level.
- To demonstrate the platform's high cell utilization, molecular measurement accuracy, and sensitivity.
Main Methods:
- Hydrodynamic differential flow resistance for improved cell utilization (>95%).
- Integrated pump/valve system for complete removal of cell-free components, ensuring accurate protein detection (R=0.96).
- Picoliter reaction chambers for sensitive detection of both mRNA transcripts and protein copies.
Main Results:
- Multi-Paired-seq achieved high cell utilization and accurate molecular measurements.
- The platform identified three distinct breast cancer cell type clusters based on multimodal data.
- Expression correlations between mRNAs and proteins were quantified under various conditions.
Conclusions:
- Multi-Paired-seq offers a powerful new tool for multimodal single-cell analysis.
- The platform facilitates comprehensive studies in cell biology, developmental biology, drug discovery, and precision medicine.
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