Optimization of Genome-wide CRISPR Screens using dual-guide RNA Infection with Cas9 Electroporation (DICE)
Stender, J. D.; Purman, C.; Lu, C.; Modi, A.; Vijaykumar, V.; Flister, M.; den Hollander, A.; Kadri, S.
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Single-guide RNA lentiviral infection with Cas9 protein electroporation (SLICE) enables CRISPR screening in primary cell types that require transient Cas9 expression yet is limited by scalability and robustness. Here, we introduce dual-guide RNA infection with Cas9 electroporation (DICE), which expresses two guides from the same lentiviral construct that target the same gene. In genome-wide screens, DICE outperformed SLICE in defining essential genes and modulators of PD-L1 expression in IFN gamma activated THP1 cells. Collectively, these data demonstrate that DICE can be utilized for reduced-scale CRISPR screens in cell types with transient Cas9 protein expression without sacrificing screening quality.
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