Introduction:　Natural　killer　cells　can　attack　cancer　cells　without　prior　sensitization,　but　their　clinical　benefit　is　limited　owing　to　their　poor　selectivity　that　is　caused　by　the　lack　of　specific　receptors　to　target　tumor　cells.　In　this　study,　we　aimed　to　endow　NK　cells　with　the　ability　to　specifically　target　glypican-3+　tumor　cells　without　producing　cell　damage　or　genetic　alterations,　and　further　evaluated　their　therapeutic　efficiency.　Methods:　NK　cells　were　modified　with　a　Gpc3　DNA　aptamer　on　the　cell　surface　via　metabolic　glycoengineering　to　endow　NK　cells　with　specific　targeting　ability.　Then,　the　G-NK　cells　were　evaluated　for　their　specific　targeting　properties,　cytotoxicity　and　secretion　of　cytokines　in　vitro.　Finally,　we　investigated　the　therapeutic　efficiency　of　G-NK　cells　against　glypican-3+　tumor　cells　in　vivo.　Results:　Compared　with　NK　cells　modified　with　a　random　aptamer　mutation　and　unmodified　NK　cells,　G-NK　cells　induced　significant　apoptosis/necrosis　of　GPC3+　tumor　cells　and　secreted　cytokines　to　preserve　the　intense　cytotoxic　activities.　Moreover,　G-NK　cells　significantly　suppressed　tumor　growth　in　HepG2　tumor-bearing　mice　due　to　the　enhanced　enrichment　of　G-NK　cells　at　the　tumor　site.　Conclusions:　The　proposed　strategy　endows　NK　cells　with　a　tumor-specific　targeting　ability　to　enhance　adoptive　therapeutic　efficiency　in　GPC3+　hepatocellular　carcinoma.　©　2023,　Springer　Science+Business　Media,　LLC.