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InTraSeq: A Multimodal Assay that Uncovers New Single-Cell Biology and Regulatory Mechanisms
Sean Beausoleil1, Majd Ariss1, Linglin Huang2,3
1Cell Signaling Technology, Inc., Danvers, Massachusetts, USA.
Research Square
|December 23, 2024
Summary
New Intracellular Transcriptomic and Protein Sequencing (InTraSeq) technology allows simultaneous measurement of RNA and protein in single cells. This multimodal approach provides a comprehensive view of cellular function, revealing novel subtypes and regulatory mechanisms.
Area of Science:
- Cell Biology
- Genomics
- Immunology
Background:
- Single-cell RNA sequencing (scRNA-seq) offers transcriptomic insights but provides an incomplete view of cellular function.
- Multimodal scRNA-seq assays are emerging to overcome this limitation by measuring both RNA and protein simultaneously.
- Existing methods may not capture the full spectrum of intracellular protein dynamics, including post-translational modifications.
Purpose of the Study:
- To introduce Intracellular Transcriptomic and Protein Sequencing (InTraSeq), a novel multimodal scRNA-seq technology.
- To demonstrate InTraSeq's capability for comprehensive cellular profiling by measuring mRNA, surface, cytoplasmic, and nuclear proteins.
- To apply InTraSeq to elucidate complex cellular states and regulatory mechanisms during Th17 cell differentiation.
Main Methods:
- Development of InTraSeq, a multimodal scRNA-seq assay utilizing oligo-barcoded antibodies.
- Concurrent measurement of mRNA and protein expression (surface, cytoplasmic, nuclear) in individual cells.
- Application of InTraSeq to profile over 85,000 cells during Th17 cell differentiation.
Main Results:
- InTraSeq successfully captured simultaneous RNA and protein expression profiles at high resolution.
- The study revealed novel insights into Th17 cell differentiation, identifying key regulatory factors and target genes.
- Simultaneous measurement of mRNA, intracellular proteins, and post-translational modifications provided a comprehensive understanding of cellular processes.
Conclusions:
- InTraSeq offers a powerful and comprehensive approach to studying cellular function by integrating transcriptomic and proteomic data.
- This technology enables the identification of novel cellular subtypes and states previously obscured by RNA-only analyses.
- InTraSeq advances the understanding of cellular heterogeneity and regulatory mechanisms, particularly in complex biological processes like immune cell differentiation.
