1 Citation
Additional file 1: Sup Figure 1. Generation of Gria2tm1BViss mice and GluA2 Q/R site editing efficiency analysis. (a) Schematic representation of the GluA2 WT allele, the targeted GluA2G/G/neo allele and the targeted GluA2G/G allele, after the removal of the floxed neo cassette by Cre-mediated recombination in ES cells. Exons 10, 11 and 12 are shown (black boxes). Black arrows indicate loxP sites. The position of the adenosine to guanine mutation is indicated in red. (b) DNA sequencing of WT and Gria2tm1BViss mice confirmed the single adenosine to guanine mutation in homozygous mice. (c) Genotype analysis of WT, GluA2G/- and GluA2G/G mice by PCR shows a band at 200 bp in WT, two bands at 200 bp and 250 bp in heterozygous mice and a single band at 250bp in homozygous mice. (d) BbvI digestion assay. Schematic representation of the GluA2 mRNA Bbv1 digestion assay shows 2 bands produced for edited GluA2 templates (225bp and 68bp) and 3 bands for unedited GluA2 template (144 bp, 81bp and 68 bp). Representative image and quantification of Bbv1 digestions revealed GluA2G/G mice exhibit 0% unedited GluA2, whereas WT animals exhibit 0.44% unedited GluA2 in the hippocampus (n = 3/genotype). Each value represents the mean ± the SD.