Establishment of a TLR3 homozygous knockout human induced pluripotent stem cell line using CRISPR/Cas9

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.