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%.

中文翻译：

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基于 CRISPR 的单细胞分子筛选设计

几个研究小组最近将 CRISPR 扰动和单细胞 RNA 测序结合起来进行汇总遗传筛选。我们证明，由于慢病毒模板转换，这些研究的载体设计很容易受到引导RNA-条形码关联约50%交换的影响。我们优化了已发布的替代方案 CROP-seq，其中向导 RNA 也充当条形码，并在此确认该策略执行稳健，并将向导分配给细胞的比率提高了一倍，达到 94%。