• ASCT2/SLC1A5 encodes a transmembrane transporter, which mainly transports glutamine and some macromolecular neutral amino acids. ASCT2 is overexpressed in many tumor cells, and high expression of ASCT2 is positively correlated with poor prognosis of tumor patients.
• Mouse Slc1a5 mRNA was only detectable in wild-type mice. Human SLC1A5 mRNA was exclusively detectable in homozygous B-hASCT2 mice but not in wild-type mice. Human ASCT2 were expressed in the liver, brain, colon, and uterus from B-hASCT2 mice.
• The exons 1-9 of mouse Slc1a5 gene that encodes the full-length protein was replaced by human SLC1A5 exons 1-8 in B-hASCT2 mice. The promoter and 5’UTR region of the mouse gene were also retained, while the 3’UTR region was replaced by the human counterpart. The human SLC1A5 expression was driven by endogenous mouse Slc1a5 promoter, while mouse Slc1a5 gene transcription and translation will be disrupted.
• This product can be used for the study of the drug mechanism or safety evaluation targeting ASCT2.

Gene targeting strategy for B-hASCT2 mice. The exons 1-9 of mouse Slc1a5 gene that encodes the full-length protein was replaced by human SLC1A5 exons 1-8 in B-hASCT2 mice. The promoter and 5’UTR region of the mouse gene were also retained, while the 3’UTR region was replaced by the human counterpart. The human SLC1A5 expression was driven by endogenous mouse Slc1a5 promoter, while mouse Slc1a5 gene transcription and translation will be disrupted.

Strain specific analysis of SLC1A5 mRNA expression in wild-type C57BL/6 mice and B-hASCT2 mice by RT-PCR. Brain RNA were isolated from wild-type C57BL/6 mice (+/+) and homozygous B-hASCT2 mice (H/H), then cDNA libraries were synthesized by reverse transcription, followed by PCR with mouse or human SLC1A5 primers. Mouse Slc1a5 mRNA was only detectable in wild-type mice. Human SLC1A5 mRNA was exclusively detectable in homozygous B-hASCT2 mice but not in wild-type mice.

Immunohistochemical analysis of ASCT2 protein expression in B-hASCT2 mice. Tissues were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hASCT2 mice (2 females, 20-weeks-old) for IHC analysis. Tissues were stained with an anti-human ASCT2 antibody (abcam, ab237704). The results demonstrated that human ASCT2 were expressed in the liver, brain, colon, and uterus from B-hASCT2 mice. The plus sign (+) denotes positive staining, while the minus sign (-) denotes negative staining. Positive signals are indicated by arrows in the figure.