Fig. 3.

Fcwf-4 IRN cells express an mRNA predicted to generate a truncated, null-mutant IFNαR protein. A) Deduced nucleotide and amino acid sequences of the IFNαR2 region targeted by Crispr/Cas technology. Single-guide RNA sequence target (underlined); protospacer-adjacent motif (PAM) (yellow); STOP codon and asterisk (red) indicate early termination of translation of the IFNαR2 protein in IRN cells. B) Feline IFNαR2 exon 1 expression determined from total RNAs collected from Fcwf-4 CU and Fcwf-4 IRN confluent monolayers. mRNA expression normalized to β-actin and presented as average 2^−ΔCt expression values. Data represent two independent experiments in triplicate. Mean ± SD analyzed by unpaired t-tests. **P < 0.01. C) Melt curves and maximum melt temperatures of IFNαR2 amplicons produced during qPCR of RNAs obtained from Fcwf-4 CU or Fcwf-4 IRN cells. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)