Bacteriotherapy is one of the important direction in the field of cancer therapy. The increasing development of synthetic biology provides more diversified, more effective and safer therapeutic strategies for bacterial anti-cancer therapy. In this review, the excellent clinical examples in the field of bacterial anti-cancer therapy are summarized, the current research progress and bottleneck problems are introduced in detail. The application of synthetic biology in promoting the clinical development of live bacterial therapeutics is also discussed. Moreover, the important role and the future development trend of synthetic biology are prospected in view of the effectiveness and biosafety of clinical live bacterial therapeutics.