Primary cells are often challenging to edit and transfect, limiting long-term viabilities, editing efficiencies, and downstream functional assays, despite their tremendous value in translational research. This protocol, developed in collaboration with Lonza, describes step-by-step instructions on editing resting primary human CD4+ T cells using Synthego’s synthetic sgRNA and Lonza 4D-Nucleofector®.

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 the step-by-step process of transfecting and culturing resting human primary CD4+ T cells under optimal conditions.

  • Required transfection reagents and equipment
  • Detailed transfection protocols, including steps on how to use the Lonza 4D-Nucleofector®
  • Data that demonstrate editing efficiency in resting human primary CD4+ T cells