{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T22:05:10Z","timestamp":1790460310843,"version":"4.1.0"},"reference-count":32,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2022,6,30]],"date-time":"2022-06-30T00:00:00Z","timestamp":1656547200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["2020YFA0908800"],"award-info":[{"award-number":["2020YFA0908800"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["81871376"],"award-info":[{"award-number":["81871376"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["82071935"],"award-info":[{"award-number":["82071935"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["81901771"],"award-info":[{"award-number":["81901771"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["JCYJ20190812171820731"],"award-info":[{"award-number":["JCYJ20190812171820731"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100012166","name":"National Key Research and Development Program of China","doi-asserted-by":"publisher","award":["JCYJ20190807144209381"],"award-info":[{"award-number":["JCYJ20190807144209381"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National Natural Science Foundation of China","award":["2020YFA0908800"],"award-info":[{"award-number":["2020YFA0908800"]}]},{"name":"National Natural Science Foundation of China","award":["81871376"],"award-info":[{"award-number":["81871376"]}]},{"name":"National Natural Science Foundation of China","award":["82071935"],"award-info":[{"award-number":["82071935"]}]},{"name":"National Natural Science Foundation of China","award":["81901771"],"award-info":[{"award-number":["81901771"]}]},{"name":"National Natural Science Foundation of China","award":["JCYJ20190812171820731"],"award-info":[{"award-number":["JCYJ20190812171820731"]}]},{"name":"National Natural Science Foundation of China","award":["JCYJ20190807144209381"],"award-info":[{"award-number":["JCYJ20190807144209381"]}]},{"name":"Guangdong Innovation Platform of Translational Research for Cerebrovascular Diseases","award":["2020YFA0908800"],"award-info":[{"award-number":["2020YFA0908800"]}]},{"name":"Guangdong Innovation Platform of Translational Research for Cerebrovascular Diseases","award":["81871376"],"award-info":[{"award-number":["81871376"]}]},{"name":"Guangdong Innovation Platform of Translational Research for Cerebrovascular Diseases","award":["82071935"],"award-info":[{"award-number":["82071935"]}]},{"name":"Guangdong Innovation Platform of Translational Research for Cerebrovascular Diseases","award":["81901771"],"award-info":[{"award-number":["81901771"]}]},{"name":"Guangdong Innovation Platform of Translational Research for Cerebrovascular Diseases","award":["JCYJ20190812171820731"],"award-info":[{"award-number":["JCYJ20190812171820731"]}]},{"name":"Guangdong Innovation Platform of Translational Research for Cerebrovascular Diseases","award":["JCYJ20190807144209381"],"award-info":[{"award-number":["JCYJ20190807144209381"]}]},{"name":"Science, Technology and Innovation Commission of Shenzhen Municipality","award":["2020YFA0908800"],"award-info":[{"award-number":["2020YFA0908800"]}]},{"name":"Science, Technology and Innovation Commission of Shenzhen Municipality","award":["81871376"],"award-info":[{"award-number":["81871376"]}]},{"name":"Science, Technology and Innovation Commission of Shenzhen Municipality","award":["82071935"],"award-info":[{"award-number":["82071935"]}]},{"name":"Science, Technology and Innovation Commission of Shenzhen Municipality","award":["81901771"],"award-info":[{"award-number":["81901771"]}]},{"name":"Science, Technology and Innovation Commission of Shenzhen Municipality","award":["JCYJ20190812171820731"],"award-info":[{"award-number":["JCYJ20190812171820731"]}]},{"name":"Science, Technology and Innovation Commission of Shenzhen Municipality","award":["JCYJ20190807144209381"],"award-info":[{"award-number":["JCYJ20190807144209381"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>The epithelial-mesenchymal transition (EMT), a process in which epithelial cells undergo a series of biochemical changes to acquire a mesenchymal phenotype, has been linked to tumor metastasis. Here, we present a novel strategy for knocking out the EMT-related Cdh2 gene, which encodes N-cadherin through CRISPR\/Cas9-mediated gene editing by an ultrasound combined with biosynthetic nanobubbles (Gas Vesicles, GVs). Polyethyleneimine were employed as a gene delivery vector to deliver sgRNA into 4T1 cells that stably express the Cas9 protein, resulting in the stable Cdh2 gene- knockout cell lines. The Western blotting assay confirmed the absence of an N-cadherin protein in these Cdh2 gene-knockout 4T1 cell lines. Significantly reduced tumor cell migration was observed in the Cdh2 gene-knockout 4T1 cells in comparison with the wild-type cells. Our study demonstrated that an ultrasound combined with GVs could effectively mediate CRISPR\/Cas9 gene editing of a Cdh2 gene to inhibit tumor invasion and metastasis.<\/jats:p>","DOI":"10.3390\/pharmaceutics14071382","type":"journal-article","created":{"date-parts":[[2022,7,1]],"date-time":"2022-07-01T01:40:36Z","timestamp":1656639636000},"page":"1382","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Biosynthetic Nanobubble-Mediated CRISPR\/Cas9 Gene Editing of Cdh2 Inhibits Breast Cancer Metastasis"],"prefix":"10.3390","volume":"14","author":[{"given":"Ruru","family":"Gao","sequence":"first","affiliation":[{"name":"Department of Medical Ultrasound, The Second Affiliated Hospital, School of Medicine, South China University of Technology, Guangzhou 510180, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qiong","family":"Luo","sequence":"additional","affiliation":[{"name":"Department of Medical Ultrasound, The Second Affiliated Hospital, School of Medicine, South China University of Technology, Guangzhou 510180, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Gastrointestinal Surgery, Shenzhen People\u2019s Hospital (The Second Clinical Medical College, Jinan University), Shenzhen 518020, China"},{"name":"Department of Gastrointestinal Surgery, Shenzhen People\u2019s Hospital (The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen 518020, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8041-0373","authenticated-orcid":false,"given":"Liming","family":"Song","sequence":"additional","affiliation":[{"name":"Department of Orthopedics, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junnan","family":"Cai","sequence":"additional","affiliation":[{"name":"SXUltrasonic (Shenzhen) Ltd., Shenzhen 518000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Xiong","sequence":"additional","affiliation":[{"name":"Department of Medical Ultrasound, The Second Affiliated Hospital, School of Medicine, South China University of Technology, Guangzhou 510180, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fei","family":"Yan","sequence":"additional","affiliation":[{"name":"Center for Cell and Gene Circuit Design, CAS Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianhua","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Medical Ultrasound, The Second Affiliated Hospital, School of Medicine, South China University of Technology, Guangzhou 510180, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,6,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"129","DOI":"10.21147\/j.issn.1000-9604.2020.02.01","article-title":"Cancer burden and trends in China: A review and comparison with Japan and South Korea","volume":"32","author":"Sun","year":"2020","journal-title":"Chin. J. Cancer Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"7","DOI":"10.3322\/caac.21654","article-title":"Cancer Statistics, 2021","volume":"71","author":"Siegel","year":"2021","journal-title":"CA Cancer J. Clin."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1699","DOI":"10.3892\/ijo.2014.2549","article-title":"The cytoplasmic domain of N-cadherin modulates MMP9 induction in oral squamous carcinoma cells","volume":"45","author":"Walker","year":"2014","journal-title":"Int. J. Oncol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1016\/j.cell.2015.08.063","article-title":"CDK7-dependent transcriptional addiction in triple-negative breast cancer","volume":"163","author":"Wang","year":"2015","journal-title":"Cell"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"4116","DOI":"10.1158\/0008-5472.CAN-08-3441","article-title":"Characterization of a naturally occurring breast cancer subset enriched in epithelial-to-mesenchymal transition and stem cell characteristics","volume":"69","author":"Hennessy","year":"2009","journal-title":"Cancer Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1146\/annurev-pathol-020117-043854","article-title":"Epithelial Mesenchymal transition in tumor metastasis","volume":"13","author":"Mittal","year":"2018","journal-title":"Annu. Rev. Pathol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"819","DOI":"10.1126\/science.1231143","article-title":"Multiplex genome engineering using CRISPR\/Cas systems","volume":"339","author":"Cong","year":"2013","journal-title":"Science"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"823","DOI":"10.1126\/science.1232033","article-title":"RNA-guided human genome engineering via Cas9","volume":"339","author":"Mali","year":"2013","journal-title":"Science"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1124\/jpet.119.257287","article-title":"CRISPR\/Cas System for genome editing: Progress and prospects as a therapeutic tool","volume":"370","author":"Sahel","year":"2019","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"7685","DOI":"10.3892\/mmr.2017.7510","article-title":"Efficient gene therapy with a combination of ultrasoundtargeted microbubble destruction and PEI\/DNA\/NLS complexes","volume":"16","author":"Wang","year":"2017","journal-title":"Mol. Med. Rep."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"976","DOI":"10.1002\/jbm.a.37346","article-title":"Polyethyleneimine-anchored liposomes as scavengers for improving the efficiency of protein-bound uremic toxin clearance during dialysis","volume":"110","author":"Shen","year":"2022","journal-title":"J. Biomed. Mater. Res. A"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"51","DOI":"10.7150\/thno.16074","article-title":"Localized delivery of shRNA against PHD2 protects the heart from acute myocardial infarction through ultrasound-targeted cationic microbubble destruction","volume":"7","author":"Zhang","year":"2017","journal-title":"Theranostics"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2534","DOI":"10.1002\/cam4.840","article-title":"Evaluation of in vivo antitumor effects of low-frequency ultrasound-mediated miRNA-133a microbubble delivery in breast cancer","volume":"5","author":"Ji","year":"2016","journal-title":"Cancer Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1016\/j.jconrel.2011.08.026","article-title":"Non-viral gene therapy for neurological diseases, with an emphasis on targeted gene delivery","volume":"157","author":"Rogers","year":"2012","journal-title":"J. Control. Release"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1016\/j.gendis.2016.08.001","article-title":"Ultrasound-targeted microbubble destruction in gene therapy: A new tool to cure human diseases","volume":"4","author":"Wu","year":"2017","journal-title":"Genes Dis."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"705","DOI":"10.1038\/nrmicro2834","article-title":"Distribution, formation and regulation of gas vesicles","volume":"10","author":"Pfeifer","year":"2012","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"385","DOI":"10.3390\/life5010385","article-title":"Haloarchaea and the formation of gas vesicles","volume":"5","author":"Pfeifer","year":"2015","journal-title":"Life"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1038\/nnano.2014.32","article-title":"Biogenic gas nanostructures as ultrasonic molecular reporters","volume":"9","author":"Shapiro","year":"2014","journal-title":"Nat. Nanotechnol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1793","DOI":"10.1007\/s10529-017-2434-y","article-title":"Haloarchaea: Worth exploring for their biotechnological potential","volume":"39","author":"Singh","year":"2017","journal-title":"Biotechnol. Lett."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"10437","DOI":"10.1073\/pnas.1512503112","article-title":"Generation of knock-in primary human T cells using Cas9 ribonucleoproteins","volume":"112","author":"Schumann","year":"2015","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1240","DOI":"10.1056\/NEJMoa1817426","article-title":"CRISPR-edited stem cells in a patient with HIV and acute lymphocytic leukemia","volume":"381","author":"Xu","year":"2019","journal-title":"N. Engl. J. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"252","DOI":"10.1056\/NEJMoa2031054","article-title":"CRISPR-Cas9 Gene Editing for Sickle Cell Disease and beta-Thalassemia","volume":"384","author":"Frangoul","year":"2021","journal-title":"N. Engl. J. Med."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1038\/s41591-019-0738-2","article-title":"Somatic gene editing ameliorates skeletal and cardiac muscle failure in pig and human models of Duchenne muscular dystrophy","volume":"26","author":"Moretti","year":"2020","journal-title":"Nat. Med."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"6065","DOI":"10.1038\/s41467-020-19821-7","article-title":"CRISPR based editing of SIV proviral DNA in ART treated non-human primates","volume":"11","author":"Mancuso","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1073","DOI":"10.1038\/mt.2008.76","article-title":"Analysis of AAV serotypes 1\u20139 mediated gene expression and tropism in mice after systemic injection","volume":"16","author":"Zincarelli","year":"2008","journal-title":"Mol. Ther."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"3728","DOI":"10.1038\/ncomms4728","article-title":"Targeted genomic rearrangements using CRISPR\/Cas technology","volume":"5","author":"Choi","year":"2014","journal-title":"Nat. Commun."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1121","DOI":"10.1517\/17425247.5.10.1121","article-title":"Ultrasound targeted microbubble destruction for drug and gene delivery","volume":"5","author":"Mayer","year":"2008","journal-title":"Expert Opin. Drug Deliv."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"525","DOI":"10.2174\/0929867327666200212100257","article-title":"Applications of micro\/nanotechnology in ultrasound-based drug delivery and therapy for tumor","volume":"28","author":"Sun","year":"2021","journal-title":"Curr. Med. Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1038\/mt.2012.185","article-title":"The possible \u201cproton sponge\u201d effect of polyethylenimine (PEI) does not include change in lysosomal pH","volume":"21","author":"Benjaminsen","year":"2013","journal-title":"Mol. Ther."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2199","DOI":"10.1177\/0300060519840890","article-title":"Ultrasound microbubble-mediated CRISPR\/Cas9 knockout of C-erbB-2 in HEC-1A cells","volume":"47","author":"Cai","year":"2019","journal-title":"J. Int. Med. Res."},{"key":"ref_31","first-page":"583","article-title":"New biomarker for lung cancer\u2014Focus on circSETD3","volume":"35","author":"Tao","year":"2021","journal-title":"J. Biol. Regul. Homeost. Agents"},{"key":"ref_32","first-page":"1177","article-title":"Downregulation of NPTX1 induces cell cycle progression through Wnt\u03b2-catenin signaling in breast cancer","volume":"35","author":"Ye","year":"2021","journal-title":"J. Biol. Regul. Homeost. Agents"}],"container-title":["Pharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4923\/14\/7\/1382\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:40:53Z","timestamp":1760139653000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4923\/14\/7\/1382"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,6,30]]},"references-count":32,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["pharmaceutics14071382"],"URL":"https:\/\/doi.org\/10.3390\/pharmaceutics14071382","relation":{},"ISSN":["1999-4923"],"issn-type":[{"value":"1999-4923","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,6,30]]}}}