On the design of CRISPR-based single-cell molecular screens.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 29457792.
- Also identified by DOI 10.1038/nmeth.4604 and PMC identifier 5882576.
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Abstract
Several groups recently coupled CRISPR perturbations and single-cell RNA-seq for pooled genetic screens. We demonstrate that vector designs of these studies are susceptible to ∼50% swapping of guide RNA-barcode associations because of lentiviral template switching. We optimized a published alternative, CROP-seq, in which the guide RNA also serves as the barcode, and here confirm that this strategy performs robustly and doubled the rate at which guides are assigned to cells to 94%.
Medical subject headings
- Clustered Regularly Interspaced Short Palindromic Repeats
- Single-Cell Analysis