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EcNΔnlpIIHCG is an engineered live bacterial therapeutic and diagnostic agent based on the *Escherichia coli* Nissle 1917 (EcN) strain. It is specifically modified with a deletion of the *nlpI* gene to enhance the biogenesis and secretion of outer membrane vesicles (OMVs). The bacterium is engineered to express a bifunctional surface display system: the ice nucleation protein (INP) fused to histone-like protein A (HlpA) for selective targeting of tumor-associated DNA and heparan sulfate, and Cytolysin A (ClyA) fused to enhanced green fluorescent protein (EGFP) for tracking and visualization. Following oral administration, EcNΔnlpIIHCG selectively colonizes the tumor microenvironment (TME), where it serves as a fluorescent tracking tool to monitor bacterial distribution and OMV-mediated delivery. It is primarily utilized as the imaging and tracking counterpart to the therapeutic strain EcNΔnlpIIHCL (ENHL), which carries lactate oxidase to neutralize acidic stress in colorectal cancer and other solid tumors.
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