Basic Information

Strain name
C57BL/6-Cd3etm1(CD3E)Cd3dtm1(CD3D)Cd3gtm1(CD3G)Tacstd2tm1(TACSTD2)/Bcgen
Common name
B-hCD3EDG/hTROP2 mice
Background
C57BL/6
Catalog number
121633
Aliases
CD3E, IMD18, T3E, TCRE, CD3e molecule; CD3D, CD3-DELTA, IMD19, T3D, CD3d molecule;  CD3G, CD3-GAMMA, IMD17, T3G, CD3g molecule); TACSTD2, EGP-1, EGP1, GA733-1, GA7331, GP50, M1S1, TROP2, tumor associated calcium signal transducer 2, Trophoblast cell-surface antigen 2

Targeting strategy

Gene targeting strategy for B-hCD3EDG/hTROP2 mice.

The exons 2-8 of mouse Cd3e gene were replaced by human CD3E exons 2-9 in B-hCD3EDG/hTROP2 mice. The exons 1-5 of mouse Cd3d and the exons 1-6 of Cd3g gene were replaced by human CD3D exons 1-5 and CD3G exons 1-7 in B-hCD3EDG/hTROP2 mice, respectively. The exon 1 of mouse Trop2 gene that encodes the full-length protein was replaced by human TROP2 exon 1 in B-hCD3EDG/hTROP2 mice.

mRNA expression analysis

Strain specific analysis of CD3D and CD3G gene expression in wild type (WT) mice and B-hCD3EDG/hTROP2 mice by RT-PCR.

Mouse Cd3d and Cd3g mRNA were detectable only in splenocytes of WT mice (+/+). Chimeric CD3D and CD3G mRNA were detectable only in homozygous B-hCD3EDG/hTROP2 mice (H/H;H/H) but not in WT mice (+/+).

Protein expression analysis

Strain specific CD3E expression analysis in homozygous B-hCD3EDG/hTROP2 mice by flow cytometry.

Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hCD3EDG/hTROP2 mice (H/H;H/H), and analyzed by flow cytometry with species-specific anti-CD3E antibody. Mouse CD3E was detectable in WT mice (+/+). Human CD3E was exclusively detectable in homozygous B-hCD3EDG/hTROP2 mice (H/H;H/H) but not in WT mice (+/+).

Strain specific TROP2 expression analysis in homozygous B-hCD3EDG/hTROP2 mice by western blot.

Skin tissue was collected from wild type (WT) mice (+/+) and homozygous B-hCD3EDG/hTROP2 mice (H/H;H/H), and analyzed by western blot with anti-TROP2 antibody. Mouse TROP2 was detectable in WT mice (+/+). Human TROP2 was detectable in homozygous B-hCD3EDG/hTROP2 mice (H/H;H/H) and B-hTROP2 MC38 cell line.

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