{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,25]],"date-time":"2026-09-25T14:27:13Z","timestamp":1790346433260,"version":"4.1.0"},"reference-count":54,"publisher":"SAGE Publications","issue":"15-16","license":[{"start":{"date-parts":[[2023,8,1]],"date-time":"2023-08-01T00:00:00Z","timestamp":1690848000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["Human Gene Therapy"],"published-print":{"date-parts":[[2023,8]]},"abstract":"<jats:p>\n                    Gene editing strategies are attractive for treating genetic pulmonary diseases such as cystic fibrosis (CF). However, challenges have included the development of safe and effective vector systems for gene editing of airway epithelia and model systems to report their efficiency and durability. The domestic ferret (\n                    <jats:italic toggle=\"yes\">Mustela putorius furo<\/jats:italic>\n                    ) has a high degree of conservation in lung cellular anatomy with humans, and has served as an excellent model for many types of lung diseases, including CF. In this study, we evaluated the efficiency of amphiphilic shuttle peptide S10 for protein delivery and gene editing using SpCas9, and AsCas12a (Cpf1) ribonucleoproteins (RNPs). These approaches were evaluated in proliferating ferret airway basal cells, polarized airway epithelia\n                    <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                    , and lungs\n                    <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                    , by accessing the editing efficiency using reporter ferrets and measuring indels at the ferret\n                    <jats:italic toggle=\"yes\">CFTR<\/jats:italic>\n                    locus. Our results demonstrate that shuttle peptides efficiently enable delivery of reporter proteins\/peptides and gene editing SpCas9 or Cpf1 RNP complexes to ferret airway epithelial cells\n                    <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                    and\n                    <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                    . We measured S10 delivery efficiency of green fluorescent protein (GFP)-nuclear localization signal (NLS) protein or SpCas9 RNP into ferret airway basal cells and fully differentiated ciliated and nonciliated epithelial cells\n                    <jats:italic toggle=\"yes\">in vitro<\/jats:italic>\n                    .\n                    <jats:italic toggle=\"yes\">In vitro<\/jats:italic>\n                    and\n                    <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                    gene editing efficiencies were determined by Cas\/LoxP-gRNA RNP-mediated conversion of a\n                    <jats:italic toggle=\"yes\">ROSA<\/jats:italic>\n                    -TG Cre recombinase reporter using transgenic primary cells and ferrets. S10\/Cas9 RNP was more effective, relative to S10\/Cpf1 RNP at gene editing of the\n                    <jats:italic toggle=\"yes\">ROSA<\/jats:italic>\n                    -TG locus. Intratracheal lung delivery of the S10 shuttle combined with GFP-NLS protein or D-Retro-Inverso (DRI)-NLS peptide demonstrated efficiencies of protein delivery that were \u223c3-fold or 14-fold greater, respectively, than the efficiency of gene editing at the\n                    <jats:italic toggle=\"yes\">ROSA<\/jats:italic>\n                    -TG locus using S10\/Cas9\/LoxP-gRNA. Cpf1 RNPs was less effective than SpCas9 at gene editing of LoxP locus. These data demonstrate the feasibility of shuttle peptide delivery of Cas RNPs to the ferret airways and the potential utility for developing\n                    <jats:italic toggle=\"yes\">ex vivo<\/jats:italic>\n                    stem cell-based and\n                    <jats:italic toggle=\"yes\">in vivo<\/jats:italic>\n                    gene editing therapies for genetic pulmonary diseases such as CF.\n                  <\/jats:p>","DOI":"10.1089\/hum.2023.016","type":"journal-article","created":{"date-parts":[[2023,6,19]],"date-time":"2023-06-19T08:24:32Z","timestamp":1687163072000},"page":"705-718","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":9,"title":["Genome Editing in Ferret Airway Epithelia Mediated by CRISPR\/Nucleases Delivered with Amphiphilic Shuttle Peptides"],"prefix":"10.1177","volume":"34","author":[{"given":"Meihui","family":"Luo","sequence":"first","affiliation":[{"name":"Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jia","family":"Ma","sequence":"additional","affiliation":[{"name":"Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA."},{"name":"College of Life Science, Ningxia University, Yinchuan, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xue","family":"Cheng","sequence":"additional","affiliation":[{"name":"Feldan Therapeutics, Qu\u00e9bec, Canada."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuang","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA."},{"name":"College of Life Science, Ningxia University, Yinchuan, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Douglas J.","family":"Bartels","sequence":"additional","affiliation":[{"name":"Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Guay","sequence":"additional","affiliation":[{"name":"Feldan Therapeutics, Qu\u00e9bec, Canada."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John F.","family":"Engelhardt","sequence":"additional","affiliation":[{"name":"Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3902-9105","authenticated-orcid":false,"given":"Xiaoming","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Anatomy and Cell Biology, University of Iowa, Iowa City, Iowa, USA."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"179","published-online":{"date-parts":[[2023,8,16]]},"reference":[{"key":"e_1_3_7_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41434-020-00197-8"},{"key":"e_1_3_7_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.lfs.2022.120375"},{"key":"e_1_3_7_4_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.biomaterials.2019.119711"},{"key":"e_1_3_7_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajps.2022.02.001"},{"key":"e_1_3_7_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jconrel.2022.01.013"},{"key":"e_1_3_7_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-019-09693-x"},{"key":"e_1_3_7_8_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.apsb.2021.05.020"},{"key":"e_1_3_7_9_2","doi-asserted-by":"publisher","DOI":"10.1080\/17425247.2022.2100342"},{"key":"e_1_3_7_10_2","doi-asserted-by":"publisher","DOI":"10.3389\/fbioe.2022.895713"},{"key":"e_1_3_7_11_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt.3471"},{"key":"e_1_3_7_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ymthe.2021.10.023"},{"key":"e_1_3_7_13_2","doi-asserted-by":"publisher","DOI":"10.1089\/hum.2022.036"},{"key":"e_1_3_7_14_2","article-title":"Tailoring hyperbranched poly(beta-amino ester) as a robust and universal platform for cytosolic protein delivery","volume":"34","author":"Liu X","year":"2022","unstructured":"Liu X, , Zhao Z, , Wu F, et al. Tailoring hyperbranched poly(beta-amino ester) as a robust and universal platform for cytosolic protein delivery. 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COL17A1 editing via homology-directed repair in junctional epidermolysis bullosa. Front Med (Lausanne), 2022; 9:976604.","journal-title":"Front Med (Lausanne)"},{"key":"e_1_3_7_24_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jconrel.2022.08.042"},{"key":"e_1_3_7_25_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.molpharmaceut.1c00770"},{"key":"e_1_3_7_26_2","doi-asserted-by":"publisher","DOI":"10.1039\/C7SC03918B"},{"key":"e_1_3_7_27_2","doi-asserted-by":"publisher","DOI":"10.1101\/gr.171264.113"},{"key":"e_1_3_7_28_2","article-title":"Membrane permeabilizing amphiphilic peptide delivers recombinant transcription factor and CRISPR-Cas9\/Cpf1 ribonucleoproteins in hard-to-modify cells","volume":"13","author":"Del'Guidice T","year":"2018","unstructured":"Del'Guidice T, , Lepetit-Stoffaes JP, , Bordeleau LJ, et al. Membrane permeabilizing amphiphilic peptide delivers recombinant transcription factor and CRISPR-Cas9\/Cpf1 ribonucleoproteins in hard-to-modify cells. 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