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Pervasive off-target activity in CRISPR-interference (CRISPRi) system [ChIP-seq]

GSE252978 Homo sapiens Genome binding/occupancy profiling by high throughput sequencing 8 samples Submitted 2024/10/11 Platform GPL18573
Summary
CRISPR/Cas system and its dCas derivatives have been widely and effectively used to alter the intended genomic target. However, the presence of off-targets due to unintended binding of sgRNAs to sequences that closely resemble the target sequence, is still a major challenge. Here, we utilized a genome-wide sgRNA library for the dCas9-KRAB CRISPRi system to investigate the presence of off-target activity and its effects on gene expression. Our study provides strong evidence that CRISPRi off-targets affect the transcriptome of a cell extensively. We also highlight that mis-matches to the target DNA are tolerated while sgRNA binding and the length of the contiguous matches in the PAM-proximal region is one of the most predictive features of binding, along with open chromatin and the DNA-RNA hybrid energy. Additionally, we provide a curated random forest model that can be used to predict the off-target activity in the dCas systems.
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Direct links to NCBI, no account and no request form: the whole study as GSE252978_RAW.tar, processed values as the series matrix, the supplementary file directory, and per-sample supplementary files for any of the 8 samples. Raw sequencing reads are also available from ENA.

Also filed as BioProject PRJNA1063362 and SRA study SRP483076. Searching any of these in the dataset finder brings you back here.

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