Nanopore sequencing of single-cell transcriptomes with scCOLOR-seq

Area of Science:

• Genomics
• Molecular Biology
• Biotechnology

Background:

• Accurate sequencing of single cells is crucial for understanding cellular heterogeneity.
• Existing long-read sequencing methods face challenges with error rates in molecular identifiers.
• Novel methods are needed to improve the fidelity of single-cell transcriptomic data.

Purpose of the Study:

• To introduce single-cell corrected long-read sequencing (scCOLOR-seq) for robust error correction.
• To enable standalone cDNA nanopore sequencing of single cells with high accuracy.
• To demonstrate the utility of scCOLOR-seq in various genomic applications.

Main Methods:

• scCOLOR-seq utilizes dimeric nucleotide building blocks for synthesizing barcodes and unique molecular identifiers.
• This approach allows for intrinsic error detection within oligonucleotide sequences.
• The method enables standalone cDNA nanopore sequencing of single cells.

Main Results:

• Demonstrated high barcode assignment accuracy using scCOLOR-seq.
• Evaluated differential isoform usage in myeloma cell lines with improved precision.
• Successfully detected fusion transcripts in a sarcoma cell line.

Conclusions:

• scCOLOR-seq provides a powerful tool for accurate error correction in single-cell long-read sequencing.
• The method significantly enhances the reliability of analyzing transcriptomic variations and detecting fusion events.
• scCOLOR-seq opens new avenues for single-cell genomics research.