ArciTect Hu CRISPR Optimization Kit, PE

This PE-conjugated kit from STEMCELL Technologies is designed to enable optimization of genome editing in human cells and can also be used as a positive control for experiments using the ArciTect CRISPR-Cas9 genome editing system. It comprises ArciTect Human B2M sgRNA, ArciTect Cas9 Nuclease, and a PE-conjugated beta-2 microglobulin (B2M) antibody. By targeting the B2M gene, which encodes a ubiquitously expressed cell-surface protein, editing efficiency can be rapidly and quantitatively assessed by flow cytometry, measured as loss of B2M expression. The kit has been tested and validated for use with the ArciTect line of genome editing products, and editing efficiency has been demonstrated in hPSCs, T cells, and CD34+ hematopoietic stem and progenitor cells (HSPCs).

The workflow involves preparing an RNP complex by combining the Cas9 nuclease with the B2M sgRNA, delivering it into human cells, and culturing for 48–96 hours to allow genome editing to occur. Editing efficiency is then assessed by collecting cells and processing them for flow cytometry using the fluorophore-conjugated B2M antibody. The included antibody is a mouse IgG1 monoclonal (Clone 35) raised against recombinant human B2M protein and is conjugated with PE (phycoerythrin) for use as a flow cytometry reagent.