Chunyu Yan, Juan Du, Yan Zhang, Lu Miao and Zhaochao Xu
{"title":"HCR对癌细胞内源性miR-21快速可视化的条件调控。","authors":"Chunyu Yan, Juan Du, Yan Zhang, Lu Miao and Zhaochao Xu","doi":"10.1039/D5AY00542F","DOIUrl":null,"url":null,"abstract":"<p >MicroRNA-21 (miR-21) is a significant tumor marker for early cancer screening. Hybridization chain reaction (HCR) as an enzyme-free signal amplification method has been used for intracellular miR-21 detection in living cells, but it is limited by unclear reaction details and prolonged detection time. In this study, by effectively regulating the HCR reaction conditions, we achieved rapid visualization of endogenous miR-21 in live cells. A pair of HCR DNA hairpins H<small><sub>1</sub></small> and H<small><sub>2</sub></small> was designed, and the effects of reaction concentration, the type of HCR products, ionic selectivity, and the reaction order of H<small><sub>1</sub></small> and H<small><sub>2</sub></small> on the HCR process were investigated. We discovered that the cascade reaction produced by HCR in cells primarily formed six distinct assemblies and was dependent on different concentrations of Na<small><sup>+</sup></small> and Mg<small><sup>2+</sup></small> salts. Importantly, it was found that the addition sequence of the two hairpin DNAs was closely related to the speed of the cascading reaction in living cells. Adding H<small><sub>1</sub></small> first and then H<small><sub>2</sub></small> resulted in stronger detection signals in a relatively short time. By elucidating these details, we carried out the rapid fluorescence colocalization detection of endogenous miR-21 in living cells.</p>","PeriodicalId":64,"journal":{"name":"Analytical Methods","volume":" 25","pages":" 5170-5175"},"PeriodicalIF":2.6000,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Conditional regulation of HCR for rapid visualization of endogenous miR-21 in cancer cells†\",\"authors\":\"Chunyu Yan, Juan Du, Yan Zhang, Lu Miao and Zhaochao Xu\",\"doi\":\"10.1039/D5AY00542F\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p >MicroRNA-21 (miR-21) is a significant tumor marker for early cancer screening. Hybridization chain reaction (HCR) as an enzyme-free signal amplification method has been used for intracellular miR-21 detection in living cells, but it is limited by unclear reaction details and prolonged detection time. In this study, by effectively regulating the HCR reaction conditions, we achieved rapid visualization of endogenous miR-21 in live cells. A pair of HCR DNA hairpins H<small><sub>1</sub></small> and H<small><sub>2</sub></small> was designed, and the effects of reaction concentration, the type of HCR products, ionic selectivity, and the reaction order of H<small><sub>1</sub></small> and H<small><sub>2</sub></small> on the HCR process were investigated. We discovered that the cascade reaction produced by HCR in cells primarily formed six distinct assemblies and was dependent on different concentrations of Na<small><sup>+</sup></small> and Mg<small><sup>2+</sup></small> salts. Importantly, it was found that the addition sequence of the two hairpin DNAs was closely related to the speed of the cascading reaction in living cells. Adding H<small><sub>1</sub></small> first and then H<small><sub>2</sub></small> resulted in stronger detection signals in a relatively short time. By elucidating these details, we carried out the rapid fluorescence colocalization detection of endogenous miR-21 in living cells.</p>\",\"PeriodicalId\":64,\"journal\":{\"name\":\"Analytical Methods\",\"volume\":\" 25\",\"pages\":\" 5170-5175\"},\"PeriodicalIF\":2.6000,\"publicationDate\":\"2025-05-28\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Analytical Methods\",\"FirstCategoryId\":\"92\",\"ListUrlMain\":\"https://pubs.rsc.org/en/content/articlelanding/2025/ay/d5ay00542f\",\"RegionNum\":3,\"RegionCategory\":\"化学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"CHEMISTRY, ANALYTICAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Analytical Methods","FirstCategoryId":"92","ListUrlMain":"https://pubs.rsc.org/en/content/articlelanding/2025/ay/d5ay00542f","RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
Conditional regulation of HCR for rapid visualization of endogenous miR-21 in cancer cells†
MicroRNA-21 (miR-21) is a significant tumor marker for early cancer screening. Hybridization chain reaction (HCR) as an enzyme-free signal amplification method has been used for intracellular miR-21 detection in living cells, but it is limited by unclear reaction details and prolonged detection time. In this study, by effectively regulating the HCR reaction conditions, we achieved rapid visualization of endogenous miR-21 in live cells. A pair of HCR DNA hairpins H1 and H2 was designed, and the effects of reaction concentration, the type of HCR products, ionic selectivity, and the reaction order of H1 and H2 on the HCR process were investigated. We discovered that the cascade reaction produced by HCR in cells primarily formed six distinct assemblies and was dependent on different concentrations of Na+ and Mg2+ salts. Importantly, it was found that the addition sequence of the two hairpin DNAs was closely related to the speed of the cascading reaction in living cells. Adding H1 first and then H2 resulted in stronger detection signals in a relatively short time. By elucidating these details, we carried out the rapid fluorescence colocalization detection of endogenous miR-21 in living cells.