Figure 1
From: Knockin of Cre Gene at Ins2 Locus Reveals No Cre Activity in Mouse Hypothalamic Neurons
Generation of Ins2-Cre knockin mice.
(a) Schematic demonstration for knockin of Cre gene at the Ins2 locus by homologous recombination. The targeting vector contains the IRES-Cre-pA and FRT-flanked Neo cassette, which was inserted between the stop codon and 3′ UTR of the 3rd exon of Ins2. After successful homologous recombination in ES cells, the recombined allele continued into the germline. The Neo cassette was removed by crossing onto Flp mice. The genotyping PCR primers (P1-P6) were indicated as their corresponding location of the genome. (b) PCR analysis of genomic DNA for the indicated ES clones. A specific band size of 5.6 kb was amplified from positive clones (9 and 13) by PCR using primers P1/P2, indicating homologous recombination at the Ins2 locus. (c) PCR genotyping revealed the identification of the mouse line with the Neo cassette deleted by Flp. In mouse #5, Neo cassette deletion was evidenced by the null PCR amplification using primers P3/P4. (d) Cre mRNA expression in different tissues from Ins2-Cre mice. PANC, pancreas; HIP, hippocampus; HYT, hypothalamus; LV, liver; KD, kidney; SM, skeletal muscle; WAT, white adipose tissue; PTU, pituitary; THY, thymus; SI, small intestine. *P < 0.001 vs pancreas. N = 4. (e) Ins1 and Ins2 mRNA expression levels in the islets were not significantly different between control and Ins2-Cre mice. P > 0.05 vs control. N = 4.
