|Method of Cryopreservation||Sperm|
|Trial IVF % Fertilization||97.30%|
|Mutation #1: RMCE Targeted Mutagenesis|
Name: pancreatic and duodenal homeobox 1; cre
|Zygosity at cryopreservation||Heterozygote|
|PCR Genotyping Protocol||Pdx1.Cre_PCR_genotyping_protocol.docx|
|Strain Type||Congenic Strain|
|Chimera/Founder Genetic Background||129S6/SvEvTac|
|Cryopreservation Strain Background (VCMR)||C57BL/6J|
|Viability and Fertility Data||
Strain Background: 93.75% C57Bl6/J at cryopreservation
Homozgyous lethal at P0.
An 8.62 kb region of this gene has been replaced by tandemly oriented Lox66 and Lox2272 sites flanking positive (puromycin) and negative (HSV-TK) selectable markers.
The Pdx1.Cre exchange vector was made by cloning Cre coding sequences immediately following the translation initiation site in the Pdx1 gene. The basal Pdx1 exchange vector contains all of the 8,610 bp of DNA from Pdx1 gene (promoter and exon 1 that is absent in the Pdx1LCA allele, Lox66 and Lox2272 sites, and a flrted (flanked by FRT) hygromycin selection cassette.
The Pdx1.cre vector was exchanged into Pdx1LCA mESCs utilizing Recombinase Mediated Cassette Exchange (RMCE) and mice were generated from the correctly exchanged mESCs. These animals were subsequently mated with Flpe mice in order to remove the HygroR cassette.