Advances in organoid technology have opened new paths for developing cancer models that more closely resemble the cell composition and pathophysiology characteristics of patients. Patient-derived tumor organoids maintain histopathology and genetic/phenotypic characteristics of original tumors after multiple passages, which can not only be used as an excellent model for screening new anticancer drugs, but also predict the clinical response of patients through drug sensitivity testing, providing a reliable basis for individualized precision treatment of cancer patients. By constructing an organoid biobank for each patient, a variety of therapeutic regimens such as targeted drugs and individual/combined chemotherapy drugs can be screened. Combined with single-cell sequencing and bulk transcriptome sequencing analysis, the sensitivity of each patient to different drugs can be predicted, which can provide a reference for clinical medication, and promote the progress of individualized precision treatment for cancer patients. Guided by engineering principles, synthetic biology offers unique tools to reconstruct spatial and dynamic signals to regulate intercellular communications. In clinical cancer treatment, synthetic biology mainly employs rational artificial design to synthesize a large number of therapeutic gene circuits, which are eventually implanted into the patient body with the assistance of vectors to correct the original circuits with defective functions and achieve the ultimate goal of disease treatment. The rapid development of synthetic biology has provided new paths and methods for developing tumor organoids, including how to engineer organoids to reconstruct spatial and dynamic signals, maintain cell homeostasis, and regulate intercellular communications. In this review, the construction process of tumor organoids and their applications in synthetic biology are summarized. The current limitations of tumor organoids in terms of construction efficiency, standardization, automation, and accuracy are discussed. Finally, we discuss the prospects of synthetic biology in engineering tumor organoids with complicated structures for specific functions.