Dataset features


Dataset type: Other
Number of samples: 22
Release date: May 8 2018
Last update date: May 8 2018
Access: Public
Chemicals: Steroids
Dataset link Inflammatory Gene Changes in Spleens from WT and Rescued GR-C3 Knockin Mice Treated with LPS

Experimental Protocol

GR-C3 knockin mice were generated by standard gene targeting procedures. In brief, ES cells were generated in which exon 2 of the NR3C1 gene was replaced by a neomycin cassette. An exchange vector was generated containing a hygromycin cassette and a mutant NR3C1 exon 2 in which the ATG start codon for GR-C3 was left intact but mutated to ATC for each of the other seven translational isoforms. The exchange vector and Cre-recombinase were transfected into the targeted ES cells by electroporation, and positive ES cells were selected using G418 (sensitive) and hygromycin (resistant). A GR-C3 positive ES cell clone was injected into C57Bl/6 blastocysts to create chimeric mice. Chimeric males were bred with C57Bl/6 females for germline transmission, and the hygromycin cassette was deleted using Flp-deleter mice. The resulting mice were maintained on a C57Bl/6 background and intercrossed to produce WT, heterozygous GR-C3 knockin, and homozygous GR-C3 knockin mice. Homozygous GR-C3 knockin mice die at birth due to respiratory distress but can be completely rescued by antenatal glucocorticoid treatment. Adult male WT and rescued homozygous GR-C3 knockin mice were injected intraperitoneally with PBS or LPS (10μg/g body weight). Mice were sacrificed 3 hours and 24 hours after LPS administration and the spleen was removed. Total RNA was isolated from the spleens (n=4 per treatment group) using TRIzol reagent according to the manufacturer's instructions.








NIEHS Microarray Core