C57BL/6JNifdc-Scn10atm1(SCN10A)Bcgen /Bcgen
• 113423Gene targeting strategy for B-hSCN10A mice. The exons 2-28 of mouse Scn10a gene that encode the whole molecule (ATG to STOP codon) were replaced by human counterparts in B-hSCN10A mice. The promoter, 5’UTR and 3’UTR region of the mouse gene are retained. The human SCN10A expression is driven by endogenous mouse Scn10a promoter, while mouse Scn10a gene transcription and translation will be disrupted.
Strain specific analysis of SCN10A mRNA expression in wild-type C57BL/6JNifdc mice and homozygous B-hSCN10A mice by RT-PCR. Dorsal root ganglia (DRG) RNA were isolated from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hSCN10A mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human SCN10A primers. Mouse Scn10a mRNA was detectable in wild-type C57BL/6JNifdc mice and homozygous B-hSCN10A mice. Human SCN10A mRNA was detectable only in homozygous B-hSCN10A mice but not in wild-type mice.
Western blot analysis of SCN10A protein expression in homozygous B-hSCN10A mice. Various tissue lysates were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hSCN10A mice (H/H), and then analyzed by western blot with anti-hSCN10A antibody (Alomone, ASC-028). 40 μg total proteins were loaded for western blotting analysis. SCN10A protein was detectable in DRG, brain, cerebellum and skin from homozygous B-hSCN10A mice and wild-type C57BL/6 mice.