A CRISPR-Cas9 delivery system for in vivo screening of genes in the immune system
A CRISPR-Cas9 delivery system for in vivo screening of genes in the immune system
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DOI：
10.1038/s41467-019-09656-2
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发表时间：
2019-04-10
影响因子：
16.6
通讯作者：
Sharpe, Arlene H.
中科院分区：
文献类型：
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作者：
LaFleur, Martin W.;Nguyen, Thao H.;Sharpe, Arlene H.
靶向免疫细胞功能的疗法在癌症和自身免疫等疾病中具有显著的临床疗效。尽管功能基因组学加速了癌症治疗靶点的发现，但其在原代免疫细胞中的应用受到限制，因为载体递送效率低下，并且可能扰乱细胞状态。在这里，我们描述CHIME：CHimeric IMmune Editing，一种CRISPR-Cas9骨髓递送系统，用于在体内快速评估先天性和适应性免疫细胞中的基因功能，而无需对这些成熟谱系进行离体操作。这种方法能够有效地删除主要免疫谱系中的目的基因，而不改变它们的发育或功能。我们使用这种方法进行体内合并遗传筛选，并确定Ptpn 2作为CD 8 + T细胞介导的对LCMV克隆13病毒感染的应答的负调节因子。这些发现表明，这种遗传平台可以通过在体内免疫细胞中进行合并筛选来实现快速靶点发现。
Therapies that target the function of immune cells have significant clinical efficacy in diseases such as cancer and autoimmunity. Although functional genomics has accelerated therapeutic target discovery in cancer, its use in primary immune cells is limited because vector delivery is inefficient and can perturb cell states. Here we describe CHIME: CHimeric IMmune Editing, a CRISPR-Cas9 bone marrow delivery system to rapidly evaluate gene function in innate and adaptive immune cells in vivo without ex vivo manipulation of these mature lineages. This approach enables efficient deletion of genes of interest in major immune lineages without altering their development or function. We use this approach to perform an in vivo pooled genetic screen and identify Ptpn2 as a negative regulator of CD8+ T cell-mediated responses to LCMV Clone 13 viral infection. These findings indicate that this genetic platform can enable rapid target discovery through pooled screening in immune cells in vivo.