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Updated: Nov 17, 2025
CRISPR-Cas9 Mediated Gene Deletion in Human Pluripotent Stem Cells Cultured Under Feeder-Free Conditions
Published on: November 1, 2024
Establishment of a TLR3 homozygous knockout human induced pluripotent stem cell line using CRISPR/Cas9
Hyeong-Jun Han1, Jung-Hyun Kim1
1Division of Intractable Diseases, Center for Biomedical Sciences, Korea National Institute of Health, Cheongju 28160, Republic of Korea; National Stem Cell Bank of Korea, Korea Institute of Health, Cheongju 28160, Republic of Korea.
Abstract:
The Toll-like receptor (TLR) family plays an important role in the recognition of pathogens, including bacteria, viruses, fungi, and parasites, followed by the activation of innate immunity. TLR3 recognizes double-stranded RNA, a form of genetic material produced by positive-strand RNA viruses and DNA viruses, and is activated by viral infection. Upon recognition, TLR3 promotes the activation of interferon regulatory factor 3 to enhance the expression and secretion of type I interferons that signal other cells to enhance their antiviral defenses. We generated biallelic mutants of the TLR3 gene using a CRISPR-Cas9 genome editing method in human induced pluripotent stem cells (hiPSCs). TLR3 homozygous-knockout hiPSCs retained normal morphology, gene expression, and in vivo differentiation potential.
