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Here we engineer a thermostable Cas9 from\n                    <jats:italic>Geobacillus stearothermophilus<\/jats:italic>\n                    (GeoCas9) to generate iGeoCas9 variants capable of &gt;100\u00d7 more genome editing of cells and organs compared with the native GeoCas9 enzyme. Furthermore, iGeoCas9 RNP\u2013LNP complexes edit a variety of cell types and induce homology-directed repair in cells receiving codelivered single-stranded DNA templates. Using tissue-selective LNP formulations, we observe genome-editing levels of 16\u201237% in the liver and lungs of reporter mice that receive single intravenous injections of iGeoCas9 RNP\u2013LNPs. In addition, iGeoCas9 RNPs complexed to biodegradable LNPs edit the disease-causing\n                    <jats:italic>SFTPC<\/jats:italic>\n                    gene in lung tissue with 19% average efficiency, representing a major improvement over genome-editing levels observed previously using viral or nonviral delivery strategies. These results show that thermostable Cas9 RNP\u2013LNP complexes can expand the therapeutic potential of genome editing.\n                  <\/jats:p>","DOI":"10.1038\/s41587-024-02437-3","type":"journal-article","created":{"date-parts":[[2024,10,16]],"date-time":"2024-10-16T06:02:21Z","timestamp":1729058541000},"page":"1445-1457","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":155,"title":["Lung and liver editing by lipid nanoparticle delivery of a stable CRISPR\u2013Cas9 ribonucleoprotein"],"prefix":"10.1038","volume":"43","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3325-3536","authenticated-orcid":false,"given":"Kai","family":"Chen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hesong","family":"Han","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-9890-048X","authenticated-orcid":false,"given":"Sheng","family":"Zhao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-2464-6111","authenticated-orcid":false,"given":"Bryant","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Boyan","family":"Yin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-3163-8803","authenticated-orcid":false,"given":"Atip","family":"Lawanprasert","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marena","family":"Trinidad","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-8256-3867","authenticated-orcid":false,"given":"Benjamin W.","family":"Burgstone","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7815-7337","authenticated-orcid":false,"given":"Niren","family":"Murthy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9161-999X","authenticated-orcid":false,"given":"Jennifer A.","family":"Doudna","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,10,16]]},"reference":[{"key":"2437_CR1","doi-asserted-by":"publisher","first-page":"816","DOI":"10.1126\/science.1225829","volume":"337","author":"M Jinek","year":"2012","unstructured":"Jinek, M. et al. 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J.A.D. is a cofounder of Azalea Therapeutics, Caribou Biosciences, Editas Medicine, Evercrisp, Scribe Therapeutics, Intellia Therapeutics and Mammoth Biosciences. J.A.D. is a scientific advisory board member at Evercrisp, Caribou Biosciences, Intellia Therapeutics, Scribe Therapeutics, Mammoth Biosciences, The Column Group and Inari. J.A.D. is also an advisor for Aditum Bio. J.A.D. is chief science advisor to Sixth Street, is a director at Altos, Johnson & Johnson and Tempus and has research projects sponsored by Apple Tree Partners. N.M. and H.H. are founders of Opus Biosciences. The other authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}]}}