Polymerization-induced self-assembly (PISA) is a robust and highly feasible route to prepare well-defined nanoparticles in a relevant high concentration reaction system. The morphology transition of novel di-block copolymer P4VP-b-PBzMA during PISA was investigated based on reversible addition-fragmentation chain transfer (RAFT) polymerization. Spherical, worm-like and vesicular aggregates were observed at different reaction periods. The chain length of P4VP block could adjust the morphologies and dimension of aggregates. In addition, hydroxypropyl methacrylate was introduced into polymerization system to figure out its impact on morphology transition during the PISA process.