Figure 6

Inhibition of miR-194 via CRISPR and its effect on inflammatory response. A CRISPR/Cas9 strategy for targeting miR-194. Two gRNA regions are shown at the top panel. miR-194 gRNA was cloned into the vector as detailed in the materials and methods section. B DNA cleavage by CRISPR/Cas9 was detected by PCR. C Expression levels of miR-194 and its target gene TLR4 of CRISPR-miR194 clones compared to IPEC-J2 cells control. D DNA sequencing confirmed deletions (green boxes) and insertions (yellow boxes) generated by CRISPR/cas9 in miR-194 sequence. E Overexpression of TLR4 and downstream genes in infected CRISPR-miR-194 clones compared to infected IPEC-J2 cells with S. Typhimurium.