Engineered miRNAs Target Caspase-1 and NLRP3

For miR specific for caspase-1 and NLRP3, target sequences were designed using Invitrogen’s RNAi Designer (http://rnaidesigner.lifetechnologies.com/rnaiexpress). The double-stranded DNA oligonucleotides were synthesized and cloned into the parental vector pcDNA6.2-GW/EmGFP-miR (Invitrogen, Carlsbad, CA). The expression cassette for miR was moved into the pENTR/CMV vector and adenovirus was made as previously reported46 (link). The miR sequences were as follows: caspase-1, top strand 5′-TGCTGAGAAAGTACTCCTTGAGAGTCGTTTTGGCCACTGACTGACGACTCTCAGAGTACTTTCT-3′ and bottom strand 5′-CCTGAGAAAGTACTCTGAGAGTCGTCAGTCAGTGGCCAAAACGACTCTCAAGGAGTACTTTCTC-3′; NLRP3, top strand 5′-TGCTGATCACAGTGGGATTCGAAACAGTTTTGGCCACTGACTGACTGTTTCGACCCACTGTGAT-3′ and bottom strand 5′-CCTGATCACAGTGGGTCGAAACAGTCAGTCAGTGGCCAAAACTGTTTCGAATCCCACTGTGATC-3′.