• Background: The AGT gene encodes a protein (pre-angiotensinogen or angiotensinogen precursor) that is expressed in the liver. This protein is cleaved by renin when blood pressure drops, forming angiotensin I. Angiotensin I is then converted into the active form, angiotensin II, by ACE. This process helps regulate blood pressure and body fluid/electrolyte balance and plays a role in conditions like essential hypertension and preeclampsia.
• Application：Efficacy validation for diseases associated with hypertension.
• Targeting strategy: The exons 1-5 of mouse Agt gene that encode the whole molecule were replaced by human counterparts in B-hAGT mice. The promoter, 5’UTR and 3’UTR region of the mouse gene were replaced by human counterparts. The exons 1-9 of mouse Ren1 gene that encode the whole molecule were replaced by human counterparts in B-hREN mice. The promoter, 5’UTR and 3’UTR region of the mouse gene were replaced by human counterparts. B-hAGT/hREN mice is obtained by crossing B-hAGT mice (113362) with B-hREN mice (113397).
• Validation: Mouse AGT was exclusively detectable in wild-type C57BL/6JNifdc. Human AGT was exclusively detectable in homozygous B-hAGT mice and homozygous B-hAGT/hREN mice. Mouse REN was exclusively detectable in wild-type C57BL/6JNifdc. Human REN was exclusively detectable in homozygous B-hAGT/hREN mice.

Gene targeting strategy for B-hAGT/hREN mice. 

The exons 1-5 of mouse Agt gene that encode the whole molecule were replaced by human counterparts in B-hAGT mice. The promoter, 5’UTR and 3’UTR region of the mouse gene were replaced by human counterparts. 

The exons 1-9 of mouse Ren1 gene that encode the whole molecule were replaced by human counterparts in B-hREN mice. The promoter, 5’UTR and 3’UTR region of the mouse gene were replaced by human counterparts.

B-hAGT/hREN mice is obtained by crossing B-hAGT mice (113362) with B-hREN mice (113397).

Strain specific AGT expression analysis in wild-type C57BL/6JNifdc, homozygous humanized B-hAGT mice and homozygous humanized B-hAGT/hREN mice by ELISA. Serum was collected from wild-type C57BL/6JNifdc (+/+) (male and female, n=3, 7-week-old), homozygous B-hAGT mice (H/H) (male, n=3, 7-week-old) and homozygous B-hAGT/hREN mice (H/H;H/H) (male and female, n=3, 7-week-old). Expression level of mouse and human AGT were analyzed by ELISA (mouse AGT ELISA kit: Abcam, ab245718; human AGT ELISA kit: Abcam, ab287170). Mouse AGT was exclusively detectable in wild-type C57BL/6JNifdc. Human AGT was exclusively detectable in homozygous B-hAGT mice and homozygous B-hAGT/hREN mice. Values are expressed as mean ± SEM.

Strain specific REN expression analysis in wild-type C57BL/6JNifdc and homozygous humanized B-hAGT/hREN mice by ELISA. Serum was collected from wild-type C57BL/6JNifdc (+/+) (male and female, n=3, 7-week-old) and homozygous B-hAGT/hREN mice (H/H;H/H) (male and female, n=3, 7-week-old). Expression level of mouse and human REN were analyzed by ELISA (mouse REN ELISA kit: Thermo Fisher, EMREN1; human REN ELISA kit: RD, DREN00). Mouse REN was exclusively detectable in wild-type C57BL/6JNifdc. Human REN was exclusively detectable in homozygous B-hAGT/hREN mice. Values are expressed as mean ± SEM.

Strain specific analysis of AGT gene expression in wild-type C57BL/6JNifdc mice and B-hAGT/hREN mice by RT-qPCR. Liver RNA were isolated from wild-type C57BL/6JNifdc mice (+/+) (male and female, n=3, 7-week-old) and homozygous B-hAGT/hREN mice (H/H;H/H) (male and female, n=3, 7-week-old). Expression level of mouse and human AGT were analyzed by qPCR. Mouse Agt was exclusively detectable in wild-type C57BL/6JNifdc. Human AGT was exclusively detectable in homozygous B-hAGT/hREN mice. Values are expressed as mean ± SEM.