SMOOTH-seq: single-cell genome sequencing of human cells on a third-generation sequencing platform

There is no effective way to detect structure variations (SVs) and extra-chromosomal circular DNAs (ecDNAs) at single-cell whole-genome level. Here, we develop a novel third-generation sequencing platform-based single-cell whole-genome sequencing (scWGS) method named SMOOTH-seq (single-molecule real-time sequencing of long fragments amplified through transposon insertion). We evaluate the method for detecting CNVs, SVs, and SNVs in human cancer cell lines and a colorectal cancer sample and show that SMOOTH-seq reliably and effectively detects SVs and ecDNAs in individual cells, but shows relatively limited accuracy in detection of CNVs and SNVs. SMOOTH-seq opens a new chapter in scWGS as it generates high fidelity reads of kilobases long.

Automated summary

There is currently no effective method to detect structure variations (SVs) and extra-chromosomal circular DNAs (ecDNAs) at single-cell whole-genome level. However, the newly developed SMOOTH-seq method shows promise in reliably and effectively detecting SVs and ecDNAs in individual cells, while having relatively limited accuracy in detecting CNVs and SNVs. SMOOTH-seq opens up new possibilities for single-cell whole-genome sequencing by providing high fidelity reads of kilobases long.