3D printing technology allows for precise control of preparing complex geometries and internal defects in printed rock analogs, while in-situ Micro-CT imaging enables real-time observation of crack behavior. The combination of these technologies offers a new research approach for studying rock crack behavior. In this study, 3D-printed rock-like specimens containing a pre-existing flaw were prepared using a gypsum powder-based 3D printer. An advanced in-situ Micro-CT system equipped with a loading device was used to quantitatively and visually investigate the crack behavior in 3D-printed specimens under uniaxial compression testing. 2D CT images obtained from in-situ compression testing at different deformations could be used to reconstruct a 3D model and visually identify the crack patterns of the extracted cracks in 3D-printed specimens. The initiation angle of cracks, volume of the pre-existing flaw, volume of newly formed cracks, and damage value with respect to strains were analyzed to quantitatively investigate crack behavior. The results indicated that within the 3D-printed specimens, tensile cracks were first initiated near the internal flaw, followed by the occurrence of shear cracks or tensile-shear mixed cracks at the flaw tips. Additionally, there was a negative linear correlation between the initiation angle of newly formed cracks and the initial flaw angle. For flaw angles in the range of 0° ≤ α ≤ 45°, a higher number of newly formed cracks were observed in the 3D-printed specimens, and the rates of increase in crack volume and damage values with strain were faster. However, for flaw angles in the range of 45° < α ≤ 90°, the results showed the opposite trend. Furthermore, through comparison with the crack behavior of natural rocks containing a single flaw, it was found that the failure modes and crack behavior of the 3D-printed specimens exhibit certain similarities with natural rocks.