Affiliation:
1. Uniklinik RWTH Aachen
2. Interdisciplinary Center for Clinical Research IZKF Uniklinik RWTH Aachen
Abstract
Abstract
Human induced pluripotent stem cells (iPS cells) are efficiently differentiated into sensory neurons. These cells express the voltage-gated sodium channel NaV1.7, which is a validated pain target. NaV1.7 deficiency leads to pain insensitivity, whereas NaV1.7 gain-of-function mutants are associated with chronic pain. Here we used CRISPR/Cas9 genome editing to generate a HA-tag NaV1.7 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 NaV1.7 surface expression in both protocols. A clinically effective NaV1.7-blocker is still missing, and we expect this iPS cell model system to be useful for drug discovery and disease modeling.
Publisher
Research Square Platform LLC