GEOapplication/xmlftp://ftp.ncbi.nlm.nih.gov/geo/series/GSE330nnn/GSE330132/primaryOK200TranscriptomicsHomo sapiensExpression profiling by high throughput sequencinghttps://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE330132GEOGSEfalseMassively Multiplexed Sequencing-Based Host Cell Reactivation Assay Profiles DNA Repair at Single-Nucleotide and -Cell Resolution [scRNA]Defects in DNA repair are clinical biomarkers for therapeutic vulnerabilities in multiple cancer types. However, the characterization of DNA repair in cancer remains bottlenecked by a lack of reliable genomic predictors and the low throughput of existing functional assays. Here, we present mmHCR-seq, a sequencing-based and massively multiplexed Host Cell Reactivation assay that enables parallel quantification of functional repair efficiency and accuracy across all major repair pathways via RNA-seq readout. We demonstrate that mmHCR-seq can detect deficiencies and measure kinetics of nine DNA repair pathways simultaneously. The single-nucleotide resolution revealed lesion-dependent kinetics for base excision repair intermediates. By integrating mmHCR-seq with single-cell RNA-seq, we mapped repair capacity in a 19-cell line pool at cellular resolution and coupled repair phenotypes to expression states. Therein, cell cycle effects were unraveled without the need for synchronization. Overall, mmHCR-seq provides a scalable, high-resolution platform for dissecting DNA repair mechanisms and identifying new repair-associated vulnerabilities in cancer.2026/08/20GSE330132GSM9718932GSM971893134281330132Homo sapiens