ArciTect™ Human CRISPR Optimization Kit

Complete kit for optimization of genome editing in human cells
概要
The ArciTect™ Human CRISPR Optimization Kit is a flow cytometry-based kit designed to enable rapid optimization of genome editing in human cells. It can also be used as a positive control for experiments using the ArciTect™ CRISPR-Cas9 genome editing system in human cells. The kit comprises ArciTect™ Human B2M sgRNA, ArciTect™ Cas9 Nuclease, and a fluorophore-conjugated beta-2 microglobulin (B2M) antibody. ArciTect™ Cas9 Nuclease must first be complexed with ArciTect™ Human B2M sgRNA, followed by delivery into human cells and subsequent culture for 48 - 96 hours. Following culture, editing efficiency can be assessed quickly by collection and processing for flow cytometry using a fluorophore-conjugated B2M antibody.

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.
Components
  • ArciTect™ Human CRISPR Optimization Kit, APC (Catalog #100-0470)
    • ArciTect™ Human B2M sgRNA, 2 nmol
    • ArciTect™ Cas9 Nuclease, 100 µg (Catalog #76002)
    • Anti-Human Beta-2 Microglobulin Antibody, Clone 35, APC, 25 tests
  • ArciTect™ Human CRISPR Optimization Kit, PE (Catalog #100-0471)
    • ArciTect™ Human B2M sgRNA, 2 nmol
    • ArciTect™ Cas9 Nuclease, 100 µg (Catalog #76002)
    • Anti-Human Beta-2 Microglobulin Antibody, Clone 35, PE, 25 tests
  • ArciTect™ Human CRISPR Optimization Kit, FITC (Catalog #100-0472)
    • ArciTect™ Human B2M sgRNA, 2 nmol
    • ArciTect™ Cas9 Nuclease, 100 µg (Catalog #76002)
    • Anti-Human Beta-2 Microglobulin Antibody, Clone 35, FITC, 25 tests
Cell Type
Hematopoietic Stem and Progenitor Cells, Intestinal Cells, Pluripotent Stem Cells, T Cells
Species
Human
Brand
ArciTect
Area of Interest
Immunology, Organoids, Stem Cell Biology
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.

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