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Sci.","IJMS"],"abstract":"<jats:p>Lipid nanoparticles (LNPs) are an effective platform for delivering therapeutic payloads, but their use for targeting specific cell populations in vivo is limited by a lack of specificity. To address this issue, we developed an innovative approach for creating targeted LNPs based on the post-insertion of protein\u2013lipid conjugates. By expanding the genetic code, a single p-azido-L-phenylalanine residue was specifically incorporated into a DARPin against murine CD8, enabling copper-free click-chemistry with dibenzocyclooctyne (DBCO) to functionalize the particle surface with DARPin conjugates. DARPin-LNP characterization demonstrated preservation of particle size and acquisition of a characteristic negative charge after DARPin insertion, confirming successful post-insertion. In vitro experiments using primary murine splenic T-cells showed that targeted LNPs selectively transfected CD8+ T-lymphocytes. These data indicate that the developed platform enables selective in vitro delivery of nucleic acids to cytotoxic lymphocytes, highlighting its potential relevance for designing safer immunotherapeutic strategies.<\/jats:p>","DOI":"10.3390\/ijms27177755","type":"journal-article","created":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T09:48:36Z","timestamp":1788256116000},"page":"7755","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Genetic Code Expansion Enables Oriented, Site-Specific Conjugation of DARPins to Lipid Nanoparticles for Selective mRNA Delivery to CD8+ T Cells"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-6063-4079","authenticated-orcid":false,"given":"Anastasiia","family":"Dakhnevich","sequence":"first","affiliation":[{"name":"Biotechnology Department, Sirius University of Science and Technology, 354349 Sirius, 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