Supplementary Figure 1: Restriction fragment length polymorphism (RFLP) analysis of mouse fibroblasts in which Ern1 was targeted.
From: The SKIV2L RNA exosome limits activation of the RIG-I-like receptors
(a) MEFs immortalized with SV40 Large T antigen were transduced with lentiviruses encoding CAS9 alone, or CAS9 plus one of two guide RNAs targeting the murine Ern1 gene. Genomic DNA was isolated at the indicated times post selection of transduced cells, amplified with PCR primers flanking the targeted site, and digested with the indicated restriction enzymes. For guide RNA 1 (top), the digested wild-type DNA results in products of 191 bp and 103 bp. Targeted alleles are resistant to BsajI digestion and are ~294 bp, with a concomitant loss of the wild-type digestion products. For guide RNA 2 (bottom), the digested wild-type allele results in products of 157 bp and 96 bp. The targeted alleles are resistant to MlucI digestion and are ~253 bp. The asterisk in the bottom panel denotes a MlucI digestion product of the PCR that is outside the targeted region.
(b) A schematic view of the restriction enzyme sites within amplified genomic DNA for each guide RNA. The NHEJ product denotes insertions/deletions at the CRISPR/CAS9 target site that disrupt the restriction enzyme site.
