A universal and cost-efficient sample labeling approach for multiplexed single-cell RNA-seq based on recombinant...

Area of Science:

• Single-cell biology
• Molecular biology
• Genomics

Background:

• Single-cell transcriptomics offers deep insights into cellular diversity but is limited by high costs.
• Existing multiplexing strategies for single-cell studies are often complex or expensive.

Purpose of the Study:

• To develop a universal, cost-effective, and scalable cell membrane labeling method for single-cell transcriptomics.
• To address the limitations of current multiplexing techniques in single-cell research.

Main Methods:

• Developed HEATag (HUH-endonuclease-agglutinin tagging), combining Duck circovirus HUH endonuclease (DCV) and wheat germ agglutinin (WGA).
• Utilized DCV for sequence-specific ssDNA conjugation and WGA for robust cell labeling.
• Demonstrated compatibility with commercial and custom single-cell omics platforms.

Main Results:

• HEATag efficiently tags cell membranes with indexed single-stranded DNA (ssDNA).
• The method is robust for both fresh and fixed cells across species.
• Achieved enhanced library preparation efficiency and minimized batch effects.

Conclusions:

• HEATag offers a universal, cost-effective solution for high-throughput single-cell studies.
• This approach facilitates broader application of single-cell transcriptomics.
• Improves scalability and efficiency in single-cell omics workflows.