Restricted accessResearch articleFirst published online 2022-11
Human IFNAR2 Mutant Generated by CRISPR/Cas9-Induced Exon Skipping Upregulates a Subset of Tonic-Like Interferon-Stimulated Genes Upon IFNβ Stimulation
Type I interferons (IFN-Is) play central roles in regulating immune responses. The role of IFNAR2 in IFN-I signaling is an open question since a previous report showed that IFNβ was still functional in the absence of IFNAR2 in mice. In this study, we report that IFN-I signaling in human monocyte-derived THP1 cells absolutely depends on IFNAR2, as determined by using a knockout mutant made by CRISPR/Cas9. Additionally, we demonstrated that a 7-bp deletion mutant (Δ7) of IFNAR2 remains responsive to IFNβ stimulation and upregulates a subset of interferon-stimulated genes (s-ISGs). The s-ISGs largely overlap with tonic ISGs, which depend on the basal expression level of IFN-I. We also showed that IFN signaling in Δ7 still depends on IFNAR2. Then, we found that the 7-bp deletion in the genome results in the loss of the entire third exon (42 bp) from the mRNA and in the expression of a functionally impaired IFNAR2. These findings clarified the requirement of IFNAR2 for human IFN-I signaling and highlighted that caution should be used with CRISPR/Cas9 technology to prevent misleading interpretations caused by residual protein expression due to exon skipping or other mechanisms.
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