Inhibition of Rac1 activation effectively suppressed podocyte motility and alleviated the podocyte defects and proteinuria induced by the loss of Kindlin-2 <i>in vivo</i> Our results identify a novel Kindlin-2-RhoGDI<i>α</i>-Rac1 signaling axis that is critical for regulation of podocyte structure and function <i>in vivo</i> and provide evidence that it may serve as a useful target for therapeutic control of podocyte injury and associated glomerular diseases.