The pain target NaV1.7 is expressed late during human iPS cell differentiation into sensory neurons as determined in high resolution imaging

Human induced pluripotent stem cells (iPS cells) are efficiently differentiated into sensory neurons. These cells express the voltage-gated sodium channel Na V 1.7, which is a validated pain target. Na V 1.7 deficiency leads to pain insensitivity, whereas Na V 1.7 gain-of-function mutants are associated with chronic pain. Here we used CRISPR/Cas9 genome editing to generate a HA-tag Na V 1.7 to follow its expression. We used two differentiation protocols for generation sensory neurons: the classical small molecule approach and a directed differentiation methodology and assessed surface NaV1.7 expression by Airyscan high resolution microscopy. Our results show that maturation of at least 49 days is necessary to observe robust Na V 1.7 surface expression in both protocols. A clinically effective Na V 1.7-blocker is still missing, and we expect this iPS cell model system to be useful for drug discovery and disease modeling.