Basic Information
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Targeting strategy
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Gene targeting strategy for B-hAPOE4 mice.
The exons 2~4 of mouse Apoe gene that encode the full-length protein were replaced by human APOE4 exons 2~4 in B-hAPOE4 mice. -
Protein expression analysis in serum
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Strain specific APOE expression analysis in homozygous B-hAPOE4 mice by ELISA. Serum collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hAPOE4 mice (H/H) were analyzed by ELISA with species-specific APOE ELISA kit. Mouse APOE was detectable in wild-type mice (A). Human APOE was exclusively detectable in homozygous B-hAPOE4 mice but not in wild-type mice (B).
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Protein expression analysis in brain
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Strain specific APOE expression analysis in homozygous B-hAPOE4 mice by ELISA. Brain collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hAPOE4 mice (H/H) were extracted and APOE was measured in the brain homogenates using species-specific APOE ELISA kit and then normalized to total protein levels. Mouse APOE was detectable in wild-type mice (A). Human APOE was exclusively detectable in homozygous B-hAPOE4 mice but not in wild-type mice (B).
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mRNA expression analysis
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Strain specific analysis of APOE4 mRNA expression in wild-type C57BL/6J mice and B-hAPOE4 mice by RT-PCR. Liver RNA were isolated from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hAPOE4 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human APOE primers. Mouse Apoe mRNA was detectable only in wild-type mice. Human APOE4 mRNA was detectable only in homozygous B-hAPOE4 mice but not in wild-type mice. And the results were confirmed via Sanger sequencing.
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Summary
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mRNA expression analysis:
Mouse Apoe mRNA was detectable in liver of wild-type C57BL/6JNfidc mice. Human APOE4 mRNA was detectable in B-hAPOE4 mice and was confirmed via Sanger sequencing.Protein expression analysis:
Human APOE4 was both detectable in serum and brain from homozygous B-hAPOE4 mice, and mouse APOE was detectable only in wild-type C57BL/6JNfidc mice.
