- The exons 1-7 of mouse Il12a gene that encode the full length coding sequence were replaced by human IL12A exons 1-7 in B-hIL12A/hIL12B/hIL12RB1/hIL12RB2 mice.
- The exons 2-8 of mouse Il12b gene that encode the full length coding sequence and 3’UTR were replaced by human IL12B exons 2-8 in B-hIL12A/hIL12B/hIL12RB1/hIL12RB2 mice.
- The extracellular and transmembrane region coding sequences of human IL12RB1 gene plus the mouse Il12rb1 cytoplasmic coding sequences were inserted into the exon1 of B-hIL12A/hIL12B/hIL12RB1/hIL12RB2 mice.
- The extracellular region coding sequences of human IL12RB2 gene plus the mouse Il12rb2 transmembrane and cytoplasmic coding sequences were inserted into the exon2 of B-hIL12A/hIL12B/hIL12RB1/hIL12RB2 mice.
Protein expression analysis
Strain specific IL12p70 (IL-12A (p35) and IL-12B (p40) active heterodimer referred to as ‘p70’) expression analysis in homozygous B-hIL12A/hIL12B/hIL12RB1/hIL12RB2 mice by ELISA.
Serum was collected in wild-type (+/+) and homozygous B-hIL12A/hIL12B/hIL12RB1/hIL12RB2 mice (H/H) stimulated with anti-mCD3ε and anti-mCD28 in vivo (n=3), and analyzed by ELISA with species-specific IL12 ELISA kit. Mouse IL12p70 was detectable in wild-type mice. Human IL12 p70 was exclusively detectable in homozygous B-hIL12A/hIL12B/hIL12RB1/hIL12RB2 mice but not in wild-type mice. ND: Not detectable.