Gene editing of immune cells

A powerful tool for basic and clinical researchers is the ability to genetically modify immune cells using targeted genome engineering approaches such as CRISPR/Cas9 technology. However, inefficient delivery methods and low expression of CRISPR-Cas9 components in T cells have made the application of this technology to primary human immune cells a challenge. Using viral vectors or plasmids to apply CRISPR-Cas9 for genome editing in T cells results in low targeting efficiency and high toxicity. These issues, however, can be overcome by using electroporation to deliver Cas9 ribonucleoprotein complexes (RNPs) in tandem with single-stranded DNA templates for homology-directed repair (HDR). To help our customers circumvent some of these challenges, we share protocols developed by our R&D team for efficient editing of CD3+ Pan T cells and CD34+ cells using electroporation to deliver Cas9 RNPs and ssDNA templates.