C57BL/6-Dgat2tm1(DGAT2)Bcgen/Bcgen • 113047
Gene targeting strategy for B-hDGAT2 mice. The exons 1-8 of mouse Dgat2 gene that encode the whole molecule (ATG to STOP codon), including promoter, 5’UTR and 3’UTR were replaced by human counterparts in B-hDGAT2 mice. The human DGAT2 expression is driven by endogenous human DGAT2 promoter, while mouse Dgat2 gene transcription and translation will be disrupted.
Strain specific analysis of DGAT2 mRNA expression in wild-type C57BL/6JNifdc mice and B-hDGAT2 mice by RT-PCR. Liver RNA were isolated from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hDGAT2 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human DGAT2 primers. Mouse Dgat2 mRNA was detectable in liver of wild-type C57BL/6JNifdc mice. Human DGAT2 mRNA was detectable only in liver of homozygous B-hDGAT2 mice but not in wild-type mice.
Strain specific DGAT2 expression analysis in wild-type C57BL/6JNifdc mice and humanized B-hDGAT2 mice by Western blot. Various tissue lysates were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hDGAT2 mice (H/H), and then analyzed by western blot with anti-DGAT2 antibody (Proteintech, 17100-1-AP). DGAT2 was detectable in liver and adipose of wild-type C57BL/6 mice and homozygous B-hDGAT2 mice due to the cross-reactivity of antibody.