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Transient receptor potential canonical (TRPC) channels regulate Ca2+ influx and are activated upon stimulation by phospholipase C-generated diacylglycerol. Herein, we report that a novel TRPC3\/C6\/C7 activator, L687, can induce cellular ASO uptake. L687-induced ASO uptake was enhanced in a dose- and incubation-time-dependent manner. L687 enhanced the knockdown activity of various ASOs both in vitro and in vivo. Notably, suppression of TRPC3\/C6 by specific siRNAs reduced ASO uptake in A549 cells. Application of BAPTA-AM, a Ca2+ chelator, and SKF96365, a TRPC3\/C6 inhibitor, suppressed Ca2+ influx via TRPC3\/C6, resulting in reduced ASO uptake, thereby suggesting that Ca2+ influx via TRPC3\/C6 is critical for L687-mediated increased ASO uptake. L687 also induced dextran uptake, indicating that L687 increased endocytosis. Adding ASO to L687 resulted in endosome accumulation; however, the endosomal membrane disruptor UNC7938 facilitated endosomal escape and enhanced knockdown activity. We discovered a new function for TRPC activators regarding ASO trafficking in target cells. Our findings provide an opportunity to formulate an innovative drug delivery system for the therapeutic development of ASO.<\/jats:p>","DOI":"10.1093\/nar\/gkae245","type":"journal-article","created":{"date-parts":[[2024,4,16]],"date-time":"2024-04-16T00:40:19Z","timestamp":1713228019000},"page":"4784-4798","source":"Crossref","is-referenced-by-count":8,"title":["A novel transient receptor potential C3\/C6 selective activator induces the cellular uptake of antisense oligonucleotides"],"prefix":"10.1093","volume":"52","author":[{"given":"Hiroto","family":"Kohashi","sequence":"first","affiliation":[{"name":"Graduate School of Pharmaceutical Sciences, Osaka University , Osaka 565-0871 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryu","family":"Nagata","sequence":"additional","affiliation":[{"name":"Graduate School of Pharmaceutical Sciences, Osaka University , Osaka 565-0871 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1102-9795","authenticated-orcid":false,"given":"Yusuke","family":"Tamenori","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences, Osaka University , Osaka 565-0871 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tomorrow","family":"Amatani","sequence":"additional","affiliation":[{"name":"Graduate School of Engineering, Kyoto University , Kyoto 615-8510 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoshifumi","family":"Ueda","sequence":"additional","affiliation":[{"name":"Graduate School of Engineering, Kyoto University , Kyoto 615-8510 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yasuo","family":"Mori","sequence":"additional","affiliation":[{"name":"Graduate School of Engineering, Kyoto University , Kyoto 615-8510 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuuya","family":"Kasahara","sequence":"additional","affiliation":[{"name":"National Institutes of Biomedical Innovation, Health and Nutrition , Osaka 567-0085 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6842-6812","authenticated-orcid":false,"given":"Satoshi","family":"Obika","sequence":"additional","affiliation":[{"name":"Graduate School of Pharmaceutical Sciences, Osaka University , Osaka 565-0871 , Japan"},{"name":"Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University , Osaka 565-0871 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1859-4444","authenticated-orcid":false,"given":"Masahito","family":"Shimojo","sequence":"additional","affiliation":[{"name":"Graduate School of Pharmaceutical Sciences, Osaka University , Osaka 565-0871 , Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2024,4,16]]},"reference":[{"key":"2024052209491095900_B1","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1016\/j.cmet.2019.01.001","article-title":"RNA-targeted therapeutics","volume":"29","author":"Crooke","year":"2019","journal-title":"Cell Metab."},{"key":"2024052209491095900_B2","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1038\/s41573-021-00162-z","article-title":"Antisense technology: an overview and prospectus","volume":"20","author":"Crooke","year":"2021","journal-title":"Nat. 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