|Method of Cryopreservation||Sperm|
|Trial IVF % Fertilization||7.89%|
|Mutation #1: RMCE Targeted Mutagenesis|
Name: targeted mutation 1.2
|Zygosity at cryopreservation||Heterozygote|
|PCR Genotyping Protocol||Sox17CreER_genotyping_protocol.docx|
|Chimera/Founder Genetic Background||129S6/SvEvTac|
|Cryopreservation Strain Background (VCMR)||C57BL/6J|
|Viability and Fertility Data||
Strain Background: 87.5% C57Bl6/J at cryopreservation.
Homozygous embryonic lethal at E10.5.
Heterozygous fertile and viable.
pSox17.LCA targeting vector contains 10.288 kb 5’ arm and 4.525 kb5 3’arm. Lox66 and Lox2272 sites are inserted flanking PuTK selection marker for positive selection for targeting events with puromycin and negative selection for RMCE events with ganciclovir.
The exchange vector used for RMCE replaces Sox17 coding sequences with CreERT2. The vector also contains a Pgk-hygromycin resistance cassette flanked by tandem FRT sites, enabling excision of the cassette after germline transmission.