Protoporphyrin IX (PpIX) as natural photosensitizer derived from administration of 5-aminolevulinic acid (5-ALA) has found clinical use for photodiagnosis and photodynamic therapy of several cancers. However, broader use of 5-ALA in oncology is hampered by its charge and polarity that result in its reduced capacity for passing biological barriers and reaching the tumor tissue. Advanced drug delivery platforms are needed to improve the biodistribution of 5-ALA. Here, we report a new approach for the delivery of 5-ALA. Squalenoylation strategy was used to covalently conjugate 5-ALA to squalene, a natural precursor of cholesterol. 5-ALA-SQ nanoassemblies were formed by self-assembly in water. The nanoassemblies were monodisperse with average size of 70 nm, polydispersity index of 0.12, and ζ-potential of +36 mV. They showed good stability over several weeks. The drug loading of 5-ALA was very high at 26 %. In human prostate cancer cells PC3 and human glioblastoma cells U87MG PpIX production was monitored in vitro upon the incubation with nanoassemblies. The results showed that they were highly efficient in generating PpIX-induced fluorescence in cancer cells and demonstrate a novel nano-delivery platform for the systemic administration of 5-ALA.