Though generally mild in humans, Zika virus (ZIKV) frequently causes severe disease and congenital defects in children born to mothers infected during pregnancy (Musso et al. NEJM 2019 PMID: 31597021; Shan et al. Cell Host Microbe 2018 PMID: 30008291). Previously, chimeric candidate vaccine YF-ZIK was shown to protect mice against lethal ZIKV infection and congenital malformations after single-dose immunization (Kum et al. npj Vaccines 2018 PMID: 30564463; Kum et al. Emerg. Microbes Infect. 2020 PMID: 32116148). Here we demonstrate that YF-ZIK is safe, induces strong antiviral immunity, and protects rhesus macaques against vigorous experimental challenge. YF-ZIK triggered within 7-14 days after a single subcutaneous dose high levels of neutralizing antibodies, which could significantly be boosted by a second immunization 4 weeks later. Passively transferred immune serum protects AG129 mice from lethal ZIKV challenge, supporting neutralizing antibodies as major correlate of protection. Humoral immunity is complemented by ZIKV-specific cellular responses, characterized by a balanced Th1/Th2 cytokine production. Transcriptional analyses demonstrate the induction of a gene expression profile that is comparable to that of the original yellow fever YF17D vaccine, involving multiple pathways favoring polyvalent immunity. When YF-ZIK vaccinated macaques were challenged with a high intradermal dose of ZIKV, viral RNA remains undetectable in most animals. Immunity may be sterilizing immunity as no elevation of ZIKV neutralizing antibodies, nor seroconversion to anti-ZIKV NS1 antibodies was evident. Protection is associated with an activation of innate and adaptive immunity early after challenge. Systems analysis unveils B-cell receptor TNFRSF17 as predictor for strong antibody responses to YF-ZIK vaccination in common with YF17D (Gaucher et al. J. Exp. Med. 2008 PMID: 19047440; Querec et al. Nat. Immunol. 2009 PMID: 19029902; Fourati et al. Nat. Immunol. 2022 PMID: 36316476 ), along with GNAS and CD207 (Langerin) as novel biomarkers predicting clinical outcomes. The favorable preclinical safety, immunogenicity and efficacy of YF-ZIK justifies future evaluation in humans.