Universal Surface Biotinylation: a simple, versatile and cost-effective sample multiplexing method for single-cell...

Area of Science:

• Single-cell genomics
• Molecular biology
• Biotechnology

Background:

• Single-cell analysis allows high-throughput examination of individual cells.
• Sample multiplexing reduces costs and enhances accuracy in single-cell experiments.
• Current cell hashing methods face limitations due to the unavailability of specific cell surface markers.

Purpose of the Study:

• To develop a universal cell labeling technique independent of specific cell surface proteins.
• To overcome limitations of existing cell hashing methods for broader single-cell analysis applications.
• To improve the efficiency and accuracy of single-cell sample multiplexing.

Main Methods:

• Developed Universal Surface Biotinylation (USB) by introducing biotin into cell surface protein amine groups.
• Utilized DNA-tagged streptavidin for specific DNA 'hashtag' labeling of each cell sample.
• Validated the USB method using differentiating mouse embryonic stem cells.

Main Results:

• Achieved effective labeling across all tested cell types using the USB technique.
• Demonstrated no discernible adverse effect on transcriptome acquisition compared to conventional methods.
• Successfully applied DNA-tagged streptavidin for sample multiplexing.

Conclusions:

• Universal Surface Biotinylation (USB) offers a versatile solution for cell labeling in single-cell analysis.
• The USB method expands the applicability of cell hashing to cell types not amenable to conventional techniques.
• This approach enhances the cost-effectiveness and accuracy of large-scale single-cell studies.