概要
This kit has been tested and validated for use with the ArciTect™ line of genome editing products. By targeting the B2M gene that encodes a ubiquitously expressed cell-surface protein, editing efficiency can be rapidly and quantitatively assessed using flow cytometry methods. This technique also enables additional evaluation of editing success such as viability monitoring and/or assessment of cell type-specific marker expression.
技术资料
Document Type | 产品名称 | Catalog # | Lot # | 语言 |
---|---|---|---|---|
Product Information Sheet | ArciTect™ Human CRISPR Optimization Kits (APC, PE, FITC) | 100-0470, 100-0471, 100-0472 | All | English |
Safety Data Sheet 1 | ArciTect™ Human CRISPR Optimization Kits (APC, PE, FITC) | 100-0470, 100-0471, 100-0472 | All | English |
Safety Data Sheet 2 | ArciTect™ Human CRISPR Optimization Kits (APC, PE, FITC) | 100-0470, 100-0471, 100-0472 | All | English |
Safety Data Sheet 3 | ArciTect™ Human CRISPR Optimization Kits (APC, PE, FITC) | 100-0470 | All | English |
Safety Data Sheet 1 | ArciTect™ Human CRISPR Optimization Kits (APC, PE, FITC) | 100-0471 | All | English |
Safety Data Sheet 1 | ArciTect™ Human CRISPR Optimization Kits (APC, PE, FITC) | 100-0472 | All | English |
数据及文献
Data
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.