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Abstract In-plane anisotropic two-dimensional (2D) materials exhibit in-plane orientation-dependent properties. The anisotropic unit cell causes these materials to show lower symmetry but more diverse physical properties than in-plane isotropic 2D materials. In addition, the artificial stacking of in-plane anisotropic 2D materials can generate new phenomena that cannot be achieved in in-plane isotropic 2D materials. In this perspective we provide an overview of representative in-plane anisotropic 2D materials and their properties, such as black phosphorus, group IV monochalcogenides, group VI transition metal dichalcogenides with 1T′ and T_dphases, and rhenium dichalcogenides. In addition, we discuss recent theoretical and experimental investigations of twistronics using in-plane anisotropic 2D materials. Both in-plane anisotropic 2D materials and their twistronics hold considerable potential for advancing the field of 2D materials, particularly in the context of orientation-dependent optoelectronic devices.