During the past decade, semiconducting polymer dots (Pdots) have been prevailing in the family of fluorescent probes due to its high photon budget, excellent photo stability, and good biocompatibility. In this study, holo-Transferrin human (Tf) was utilized to covalently couple with Pdots for a highly efficient endocytosis process through transferrin receptors (TfRs) mediated internalization. As a result, the endocytosis efficiency of Tf-conjugated Pdots in HeLa cells dramatically increased as compared to that of unconjugated Pdots in the same condition. This acute increment demonstrates that holo-Transferrin molecules are of great capability for intracellular delivery of Pdots to TfRs overexpressed cells. The transportation route of Tf-conjugated Pdots is quite different from the uptake mechanism of unconjugated Pdots via nonspecific endocytic trafficking pathway. Considering the overexpression of TfRs in various cancer cells, Tf-conjugated Pdots hold potential to function as a nanocarrier for efficient drug delivery in cancer diagnostics and therapy.