Functional validation of tensin2 SH2-PTB domain by CRISPR/Cas9-mediatedgenome editing

Journal of Veterinary Medical Science

Hayato SASAKI¹  Nobuya SASAKI¹  Rieko YANOBU-TAKANASHI²  Kenta NAKANO¹  Junpei KIMURA³  Tadashi OKAMURA²  Kiyoma MARUSUGI¹ 

[1] Laboratory of Laboratory Animal Science and Medicine, School of Veterinary Medicine, Kitasato University, Towada, Aomori 034-8628, Japan;Department of Laboratory Animal Medicine, Research Institute, National Center for Global Health and Medicine, Tokyo 162�?8655, Japan;Laboratory of Anatomy, Department of Biomedical Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Hokkaido 060�?0818, Japan

关键词: CRISPR/Cas9;    glomerular sclerosis;    podocyte;    SH2-PTB domain;    tensin2;   

DOI  :  10.1292/jvms.16-0205

学科分类：兽医学

来源: Japanese Society of Veterinary Science

【 摘 要 】

References(38)Podocytes are terminally differentiated and highly specialized cells in the glomerulus,and they form a crucial component of the glomerular filtration barrier. The ICGN mouse isa model of glomerular dysfunction that shows gross morphological changes in the podocytefoot process, accompanied by proteinuria. Previously, we demonstrated that proteinuria inICR-derived glomerulonephritis mouse ICGN mice might be caused by a deletion mutation inthe tensin2 (Tns2) gene (designated Tns2nph).To test whether this mutation causes the mutant phenotype, we created knockout (KO) micecarrying a Tns2 protein deletion in the C-terminal Src homology and phosphotyrosinebinding (SH2-PTB) domains (designated Tns2ΔC) viaCRISPR/Cas9-mediated genome editing.Tns2nph/Tns2ΔC compoundheterozygotes and Tns2ΔC/Tns2ΔChomozygous KO mice displayed podocyte abnormalities and massive proteinuria similar toICGN mice, indicating that these two mutations are allelic. Further, this result suggeststhat the SH2-PTB domain of Tns2 is required for podocyte integrity. Tns2knockdown in a mouse podocyte cell line significantly enhanced actin stress fiberformation and cell migration. Thus, this study provides evidence that alteration of actinremodeling resulting from Tns2 deficiency causes morphological changes in podocytes andsubsequent proteinuria.

【 授权许可】

Unknown