Tg(CAG-EGFP,-dsRed2/RNAi:Tardbp)6Zxu

The transgenic construct contrains (from 5' to 3') the CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter, a loxP site, an enhanced green fluorescent protein sequence followed by three polyadenylation signals, a second loxP site, a dsRed2 fluorescent protein sequence, a synthetic artificial intron containing the amiR-TDP43 precursor miRNA sequence, and a polyA signal. The synthetic artificial intron utilized here was designed to contain typical intron splicing regulatory elements and was specifically placed 10 nucleotides downstream of the RFP stop codon because introns within 50 nucleotides downstream from the stop codon are found in some eukaryotic mRNAs and do not cause non-sense mediated decay (NMD). The amiR-TDP43 sequence (miR-TDP-43b; 5'-ATGCTGAACCTAAGCATAA-3') is an artificial precursor miRNA sequence encoding a short hairpin RNA specifically synthesized to target the RNA of mouse TAR DNA binding protein (TBP-43 or Tardbp). (J:207750)