{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,1]],"date-time":"2026-11-01T04:12:14Z","timestamp":1793506334149,"version":"4.5.0"},"reference-count":46,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"43","content-domain":{"domain":["www.science.org"],"crossmark-restriction":true},"short-container-title":["Sci. Adv."],"published-print":{"date-parts":[[2025,10,24]]},"abstract":"<jats:p>Lipid nanoparticles (LNPs) are a promising nonviral delivery system for gene-editing proteins, but optimal formulations remain underexplored. Unlike messenger RNA\u2013based approaches, ribonucleoprotein delivery enables immediate genome editing without relying on endogenous translation. However, intracellular delivery remains a major challenge due to protein size, charge variability, and susceptibility to denaturation and degradation. Here, we present a multistep screening platform to optimize LNP formulations for gene-editing protein delivery, focusing on in vivo T cell targeting. Through in vitro screening of a composition library, we identified top-performing candidates. In vivo screening in Ai9 mice revealed a spleen-tropic LNP formulation that preferentially targets T cells, enabling efficient gene editing in vivo. Using this LNP formulation, we achieved targeted knockout of CCR5 and PD-1 in splenic T cells, supporting potential applications in HIV resistance and cancer immunotherapy. Furthermore, a machine learning\u2013guided mechanistic study revealed key design principles for LNP-based protein delivery, highlighting unexplored opportunities for in vivo genome-editing therapies.<\/jats:p>","DOI":"10.1126\/sciadv.ady5579","type":"journal-article","created":{"date-parts":[[2025,10,22]],"date-time":"2025-10-22T17:59:42Z","timestamp":1761155982000},"update-policy":"https:\/\/doi.org\/10.34133\/aaas_crossmark","source":"Crossref","is-referenced-by-count":15,"title":["A multistep platform identifies spleen-tropic lipid nanoparticles for in vivo T cell\u2013targeted delivery of gene-editing proteins"],"prefix":"10.1126","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-1588-1772","authenticated-orcid":true,"given":"Xiaoya","family":"Lu","sequence":"first","affiliation":[{"name":"Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, USA."},{"name":"Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD, 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