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Updated: Jul 16, 2026
Establishing a High Throughput Epidermal Spheroid Culture System to Model Keratinocyte Stem Cell Plasticity
Published on: January 30, 2021
CRISPR/Cpf1-mediated knockout of FLG in human induced pluripotent stem cells generates a model for studying epidermal
E R Ntzani1, H Ramachandran2, M Papadopoulou3
1Environmental Adaptation and Cellular Resilience Laboratory, IUF - Leibniz Research Institute for Environmental Medicine, Düsseldorf, Germany; AHR Signaling & Environmentally-induced Skin Damage Laboratory, IUF - Leibniz Research Institute for Environmental Medicine, Düsseldorf, Germany.
Abstract:
Loss of filaggrin (FLG) function impairs skin barrier formation and contributes to common inflammatory skin diseases. In this study, we established a FLG knockout human induced pluripotent stem cell (iPSC) line based on KOLF2.1 J using CRISPR/Cas12a (Cpf1)-mediated genome editing. A guide RNA targeting exon 2 introduced a homozygous mutation, which was confirmed by sequencing. The edited cells maintained typical pluripotent stem cell morphology, expressed key undifferentiated markers, and retained the ability to differentiate into all three germ layers. Karyotype and copy number variation (CNV) analyses confirmed genomic stability and parental origin; the cells were free of mycoplasma. This cell line enables studies of FLG-associated skin biology and pathology.
