A great deal of research has recently been focused on Ruddlesden-Popper (RP) two-dimensional planar faults consisting of intervened [AO] monolayers in an ABO(3) perovskite framework due to the structurally peculiar shear configuration. In this work, we scrutinize the effect of elastic strain on the generation behavior of RP faults, which are electrocatalytically very active sites for the oxygen evolution reaction (OER), in (001) epitaxial LaNiO3 thin films through by using two distinct single-crystal substrates with different cubic lattice parameters. Atomic-scale direct observations reveal that RP faults can be more favorably created when tensile misfit strain is exerted. Furthermore, we demonstrate that the controlled growth of thin films show notably enhanced OER activity by the RP faults. The findings in this study highlight the impact of symmetry-breaking defect formation for better oxygen electrocatalysis in perovskite oxides.