The robust nuclear-cytoplasmic transport is vital for eukaryotes’ life cycle. Not only proteins and RNAs are transferred to their destination by this system, the important cellular activities such as mitosis are also regulated by the transport pathways via adjusting the local concentrations of some critical cargoes. tRNA is one of the most important biomacromolecules in the cell, synthesized in nucleus, and taking part in protein synthesis in cytoplasm. It is a long-held belief that tRNA functions only as a key player in protein synthesis, and tRNA nuclear-cytoplasmic transport is a unidirectional movement from nucleus to cytoplasm. However, the recent discoveries are overturning the traditional opinions. Nuclear-born tRNA can be exported out of nucleus, while cytoplasmic-residing tRNA can also be retrogradely transport into nucleus. The new concept tRNA nuclear-cytoplasmic dynamics was coined in 2008 to describe the tRNA communication between nucleus and cytoplasm, which was found able to regulate protein synthesis and cell cycle in S. cerevisiae. The latest results in this field are going to change the landscape of eukaryotic tRNA biology.