Generation of Intestinal Epithelial TTP Knockout Mice

Villin:Cre mice were obtained from Jackson Laboratories and Zfp36^fl/fl mice have been described previously^{19 (link)}. Intestinal epithelial cell-specific TTP knockout mice, termed ΔIEC, were generated by crossing Zfp36^fl/fl mice with Villin:Cre mice expressing CRE recombinase under the control of the Villin promoter. All experiments were performed using Zfp36^fl/fl littermates as controls. Mice were routinely genotyped by isolating genomic DNA from a 2 mm tail biopsy as described previously⁵⁸ . The floxed Zfp36 allele was amplified as described previously and the Cre transgene was amplified as recommended by Jackson Laboratories using the indicated primers (Table S2)^{19 (link)}. Tissue-specific knockout was confirmed by first using the MyTaq Extract-PCR Kit (Bioline) to isolate genomic DNA from 2 cm of colon tissue. Subsequently, multiplexed PCR was used to amplify either the floxed or deleted Zfp36 alleles with the oligonucleotides described previously and listed in Table S2^{19 (link)}. Unless otherwise indicated, all experiments were performed on 6-8-week-old mice. All animal breeding and experiments were approved by the Institutional Animal Care and Use Committee (IACUC) at the Pennsylvania State University College of Medicine. All experiments were performed in accordance with relevant guidelines and regulations provided by the IACUC.