Cell ReportsGenomicsPeer-reviewed journal article
Detection of deleterious on-target effects after HDR-mediated CRISPR editing
Isabel Weisheit, Joseph A. Kroeger, Rainer MalikiD, Julien KlimmtiD, Dennis Crusius, Angelika DannertiD, Martin DichgansiD, Dominik PaquetiD
Abstract
CRISPR genome editing is a promising tool for translational research but can cause undesired editing outcomes, both on-target at the edited locus and off-target at other genomic loci. We investigated the occurrence of deleterious on-target effects in human stem cells after insertion of disease-related mutations by homology-directed repair (HDR). We identified large, mono-allelic genomic deletions and loss-of-heterozygosity that escaped standard quality controls in up to 40% of edited clones. To reliably detect such events, we developed simple, low-cost and universally applicable quantitative genotyping PCR (qgPCR) as well as sequencing-based tools and suggest their usage as additional quality controls after editing. This will help to ensure the integrity of edited loci and increase the reliability of CRISPR editing.
- organism: human
- assay: CRISPR screen
- research focus: variant calling
- research focus: variant effect prediction
- approach: statistical model
- approach: experimental assay
- Citations
- 125
- FWCI
- 6.73
- Field percentile
- top 2%
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