Developing Macrophage Reporter Assays to Assess Immune Cell Responses to Nucleic Acid Therapeutics

National Center For Advancing Translational Sciences

Investigators

Abstract

During this period, the project team developed cell-based assays for detection of nucleic acid activators of immune cells. An immortalized human monocytic cell line, THP-1, was utilized to assess the capacity to engineer a luminescent reporter tag into endogenous inflammatory genes using CRISPR/Cas9. Responsiveness of the reporters to various toll-like receptor (TLR) ligands was characterized. Utilizing the knowledge gained from THP1 models, the team then pursued an iPSC derived macrophage model. Several reporters were engineered into the iPSC cells using CRISPR/Cas9. iPSC macrophages were produced and characterized using flow cytometry. The team will continue to develop this approach towards establishing a robust immune activation assay.