Extended Data Fig. 4: C865S ‘knock-in’ leads to comprised innate antiviral immunity in mice.
From: Targeting a key disulfide linkage to regulate RIG-I condensation and cytosolic RNA-sensing
a, Immunoblot (IB) analysis of the endogenous RIG-I protein levels in WT and RIG-I knockout (KO) HEK293T cells. b, Schematic diagram of Rig-IC865S/C865 knock-in strategy (upper). Rig-IC865S/C865 mice in C57BL/6N background was generated by Cyagen Biosciences Inc. by targeting of exon 16 of Rig-I using CRISPR–Cas9. Sequencing verification of the codon replacement by CRISPR–Cas9 resulting in the RIG-I C865S mutant (bottom). c, The Rig-IC865S/C865 mice were largely normal in appearance and weight and were fertile. d,e, qPCR analysis of Ifnb1, Cxcl10 or Ccl5 mRNA in wild-type and Rig-IC865S/C865 MEFs (d) and BMDMs (e) infected with SeV (top) or VSV (bottom), or transfected with 5′-ppp RNA (middle) or poly (I:C) (bottom), for indicated time periods. All results are presented relative to those of 18S; n = 3 for each experiment. Data are representative of at least three independent experiments. Mean ± s.d., statistical analysis was performed using a two-tailed Student’s t-test (d-e); **P < 0.01, ***P < 0.001, ****P < 0.0001. Exact P values, source numerical data and unprocessed blots are provided.
