Figure 4. A gene targeting strategy used to generate miR-182 KO mice. A: The pre-miR-182 locus was replaced with the Neo cassette flanked by LoxP sites (black triangles). Primers used for genotyping are shown. B: Genotyping the miR-182 KO mice. Genomic DNA isolated from wild-type (+/+), heterozygous (+/−), or homozygous KO (−/−) mice was analyzed by PCR using the primers 183–03K-F and Neo-R1 or 183–03K-F and 182-R4. C: Mature miR-182 is indeed absent in the eyes of KO mice. Total RNA was isolated from eyes of wild-type (+/+), heterozygous (+/−), and homozygous (−/−) KO mice and the expression levels of miR-96, miR-182, and miR-183 were analyzed by northern blotting. Abbreviations: long homology arm (LHA); short homology arm (SHA); neomycin resistance gene (Neo).