Chromatin assembly in eukaryote is a well-organized process in which DNA and histone are hierarchically packaged into the nucleosome and chromatin. The assembly and disassembly of chromatin decode the genetic information stored within the chromatin. Histone variant and histone chaperone are major players in regulating the chromatin assembly and virtually involve in all DNA-based processes. In this review, we discuss the recent findings that provide insights into the assembly mechanism of histone variant H2A.Z and CENP-A, and highlight the important roleshistone chaperones play in this process.