High-throughput gene expression analysis with TempO-LINC sensitively resolves complex brain, lung and kidney...

Area of Science:

• Genomics
• Molecular Biology
• Bioinformatics

Background:

• Single-cell transcriptomics is crucial for understanding cellular heterogeneity.
• Existing methods can be limited by throughput, cost, or complexity.
• A need exists for scalable and sensitive single-cell gene expression analysis.

Purpose of the Study:

• To develop and validate TempO-LINC, a novel genomics platform for high-throughput single-cell and single-nucleus transcriptomic analysis.
• To demonstrate the scalability, sensitivity, and accuracy of TempO-LINC across diverse sample types.
• To showcase TempO-LINC's utility in identifying and annotating cell populations in large-scale studies.

Main Methods:

• TempO-LINC utilizes a combinatorial indexing approach with cell-identifying molecular barcodes added to gene expression probes in fixed cells.
• The assay avoids cDNA generation, directly profiling gene expression.
• It employs over 5.3 million unique barcodes for high-resolution profiling.

Main Results:

• TempO-LINC achieved a multiplet rate below 1.1% and a cell capture rate of approximately 50%.
• The platform successfully profiled 89,722 cells, identifying over 50 unique cell populations.
• High-sensitivity gene detection was demonstrated across various sample types, including mouse lung, kidney, and brain nuclei.

Conclusions:

• TempO-LINC is a robust and scalable single-cell technology for high-throughput transcriptomic analysis.
• It offers high data quality and can be targeted for specific gene sets, reducing sequencing burden.
• This platform is well-suited for large-scale applications and studies involving thousands of samples.