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Joint single-cell profiling of Cas9 edits and transcriptomes reveals on- and off-target effects on gene expression (Superb-seq)

GSE284207 Homo sapiens Expression profiling by high throughput sequencing 4 samples Submitted 2025/02/25 Platform GPL34284Platform GPL30173Platform GPL24676
Summary
A longstanding barrier in genome engineering with CRISPR-Cas9 has been the inability to measure Cas9 edit outcomes at single-cell resolution. Here we present Superb-seq, a new technology for joint measurement of on-target and off-target Cas9 edits and transcriptomes by single-cell RNA sequencing. In contrast to Perturb-seq methods that read each cell’s guide RNA, Superb-seq directly reads Cas9 edits by leveraging T7 in situ transcription. We performed Superb-seq on 9,500 K562 cells, targeting four chromatin remodeler genes with seven guide RNAs. Superb-seq identified 11,891 edit events in 6,230 edited cells at all seven on-target sites and at 36 off-target sites. One notable off-target edit fell within the first intron of the deubiquitinase gene USP9X, which decreased its expression and perturbed the expression of downstream genes. Superb-seq uses off-the-shelf kits, standard equipment, and requires no virus, which will enable CRISPR screens in virus-intolerant cell types and functional characterization of off-target events.
Published in
Joint single-cell profiling of Cas9 edits and transcriptomes reveals widespread off-target events and effects on gene expression
Lorenzini MH, Balderson B, Sajeev K et al. · bioRxiv : the preprint server for biology 2025 · PMID 40909645 · doi:10.1101/2025.02.07.636966
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Direct links to NCBI, no account and no request form: the whole study as GSE284207_RAW.tar, processed values as the series matrix, the supplementary file directory, and per-sample supplementary files for any of the 4 samples. Raw sequencing reads are also available from ENA.

Also filed as BioProject PRJNA1197977 and SRA study SRP551283. Searching any of these in the dataset finder brings you back here.

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