• BCRP consists of 655 amino acids and possesses six transmembrane helices and one ATP­ binding site. Because BCRP is a half­-transporter, current evidence suggests that homodimerization or multimerization is required for transporter activity as illustrated.
• The exons 2~16 of ratBcrpgene were knocked out in B-Bcrp KO rats, resulting in a disruption of the Bcrp gene.
• Bcrp mRNA and protein were detectable only in wild-type SD rats, but not in B-Bcrp KO rats.

Suggestion:

• Applicable for investigating ATP-powered transport proteins in physiological drug distribution.

Gene targeting strategy for B-Bcrp KO rats.The exons 2~16 of mouseBcrpgene were knocked out in B-Bcrp KO rats, resulting in a disruption of theBcrpgene.

BcrpmRNA expression in wild-type SD rats and B-Bcrp KO rats by RT-PCR. Kidney, brain and liver RNA were isolated from wild-type SD rats (+/+) and B-Bcrp KO rats (-/-), then cDNA libraries were synthesized by reverse transcription, followed by PCR with ratBcrpprimers. RatBcrpmRNA was detectable only in wild-type SD rats, but not in B-Bcrp KO rats.