Primary cells are often difficult to transfect and manipulate, however its value in translational research is immense. In this protocol, researchers at the University of Washington, in conjunction with Synthego, describe step by step instructions on how to edit primary human CD4+ T cells by CRISPR-Cas9 using electroporation.

Although primary cells can be challenging to transfect and manipulate, Synthego’s chemically modified synthetic sgRNAs are of exceptional purity and consistently drive high editing frequencies.

This protocol describes how to deliver ribonucleoprotein (RNP) complexes that consist of purified Cas9 nuclease duplexed with chemically modified synthetic single guide RNA (sgRNA) to stimulated human primary CD4+ T cells. RNP delivery is accomplished using the Thermo Fisher Neon™ Transfection System.

This protocol provides all the necessary details to carry out your CRISPR experiment including:

  • Required transfection reagents and equipment
  • Detailed electroporation protocols including pre- and post-electroporation steps
  • Data that demonstrate editing efficiency in human primary CD4+ T cells

Download the CD4+ T Cell Neon Electroporation protocol today!