ArciTect™ Human CRISPR Optimization Kit

总览

ArciTect™ 人 CRISPR 优化试剂盒是一款基于流式细胞术的试剂盒，旨在快速优化人细胞中的基因组编辑。它也可用作在人细胞中使用 ArciTect™ CRISPR-Cas9 基因组编辑系统进行实验的阳性对照。该试剂盒包含 ArciTect™ 人 B2M sgRNA、ArciTect™ Cas9 核酸酶和荧光团偶联的 β-2 微球蛋白 (B2M) 抗体。ArciTect™ Cas9 核酸酶必须先与 ArciTect™ 人 B2M sgRNA 复合，然后递送至人细胞并培养 48 - 96 小时。培养后，可以通过收集并处理荧光团偶联的 B2M 抗体进行流式细胞术分析来快速评估编辑效率。

该试剂盒已通过测试和验证，可与 ArciTect™ 系列基因组编辑产品配合使用。通过靶向编码普遍表达的细胞表面蛋白的B2M基因，可以使用流式细胞术快速定量评估编辑效率。该技术还可以用于评估编辑成功率，例如活力监测和/或评估细胞类型特异性标记物的表达。

细胞类型
造血干/祖细胞，肠道细胞，多能干细胞，T 细胞

种属
人

应用
基因组编辑

研究领域
免疫，类器官，干细胞生物学

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实验数据

Figure 1. ArciTect™ Human CRISPR Optimization Kit Exhibits High Editing Efficiency in hPSCs, T Cells, and CD34+ Hematopoietic Stem and Progenitor Cells (HSPCs)

Cells were electroporated with RNP complexes containing sgRNA targeting B2M complexed with ArciTect™ Cas9 Nuclease protein. The cells were cultured for 72 hours following electroporation and editing efficiency was monitored by flow cytometry to detect loss of B2M expression; n = 3 biological replicates (WLS-1C iPSCs/hPSCs) or n = 3 donors (T cells and CD34+ HSPCs). No EP: non-electroporated controls.

Figure 2. Anti-Human Beta-2-Microglobulin, Clone #35 Conjugates Exhibit Equivalent Performance Across Samples to Monitor B2M Knockout Efficiency Using the ArciTect™ Human CRISPR Optimization Kit

CD34+ HSPCs isolated from two independent donors were electroporated with RNP complexes containing sgRNA targeting B2M complexed with ArciTect™ Cas9 Nuclease protein. The cells were cultured for 96 hours following electroporation and editing efficiency was monitored by flow cytometry to detect loss of B2M expression using anti-human beta-2-microglobulin, clone #35, (A, B) APC-conjugated, (C, D) FITC-conjugated, and (E, F) PE-conjugated. The histograms were derived from gated events of viable (7-AAD-negative) cells. Orange histograms: RNP-electroporated samples; grey histograms: non-electroporated control samples.