Significance EBNA1 is the only Epstein–Barr virus (EBV) latent protein responsible for viral genome maintenance and is expressed in all EBV-infected cells. Zn ²⁺ is essential for oligomerization of the functional EBNA1. We constructed an EBNA1 binding peptide with a Zn ²⁺ chelator to create an EBNA1-specific inhibitor (ZRL ₅ P ₄ ). ZRL ₅ P ₄ by itself is sufficient to reactivate EBV from its latent infection. ZRL ₅ P ₄ is able to emit unique responsive fluorescent signals once it binds with EBNA1 and a Zn ²⁺ ion. ZRL ₅ P ₄ can selectively disrupt the EBNA1 oligomerization and cause nasopharyngeal carcinoma (NPC) tumor shrinkage, possibly due to EBV lytic induction. Dicer1 seems essential for this lytic reactivation. As can been seen, EBNA1 is likely to maintain NPC cell survival by suppressing viral reactivation.