On the design of CRISPR-based single-cell molecular screens
On the design of CRISPR-based single-cell molecular screens
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DOI：
10.1038/nmeth.4604
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发表时间：
2018-04-01
期刊：
影响因子：
48
通讯作者：
Trapnell, Cole
中科院分区：
文献类型：
--
作者：
Hill, Andrew J.;McFaline-Figueroa, Jose L.;Trapnell, Cole
最近，几个研究小组将CRISPR扰动和单细胞RNA-seq结合起来，用于合并遗传筛选。我们证明，由于慢病毒模板转换，这些研究的载体设计对向导RNA-条形码关联的类似50%交换敏感。我们优化了已发表的替代方案CROP-seq，其中指导RNA也用作条形码，并且在这里证实了这种策略的稳健性，并将指导分配给细胞的比率提高了一倍，达到94%。
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 similar 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%.