Geng Qiuyu , Xing Hanxin , Cang Song , Fan Ronghua , Zheng Anqi
{"title":"利用CRISPR/Cas9灵敏检测Cu/Zn SOD mRNA","authors":"Geng Qiuyu , Xing Hanxin , Cang Song , Fan Ronghua , Zheng Anqi","doi":"10.1016/j.talo.2025.100568","DOIUrl":null,"url":null,"abstract":"<div><div>Cu/Zn SOD mRNA plays a critical role in protecting cells from oxygen toxicity by regulating the expression of the Cu/Zn SOD enzyme. In this work, we developed a biosensor, termed the Cas9-sgRNA/blocker system, for the analysis of Cu/Zn SOD mRNA. The biosensor was assembled by hybridizing a crRNA with a complementary blocker strand, which was then complexed with the Cas9 nuclease. In the presence of the target mRNA, competitive binding between the blocker strand and the mRNA restored Cas9-sgRNA cleavage activity, enabling enzymatic cleavage of adjacent fluorescent dsDNA reporters. Fluorescence measurements were performed at λex/λem = 488/525 nm, yielding a detection limit of 1.40 nmol L⁻¹. This method demonstrated excellent selectivity for Cu/Zn SOD mRNA, as evidenced by its successful application to the detection of Cu/Zn SOD mRNA in real samples.</div></div>","PeriodicalId":436,"journal":{"name":"Talanta Open","volume":"12 ","pages":"Article 100568"},"PeriodicalIF":3.7000,"publicationDate":"2025-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Sensitive detection Cu/Zn SOD mRNA with CRISPR/Cas9\",\"authors\":\"Geng Qiuyu , Xing Hanxin , Cang Song , Fan Ronghua , Zheng Anqi\",\"doi\":\"10.1016/j.talo.2025.100568\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div>Cu/Zn SOD mRNA plays a critical role in protecting cells from oxygen toxicity by regulating the expression of the Cu/Zn SOD enzyme. In this work, we developed a biosensor, termed the Cas9-sgRNA/blocker system, for the analysis of Cu/Zn SOD mRNA. The biosensor was assembled by hybridizing a crRNA with a complementary blocker strand, which was then complexed with the Cas9 nuclease. In the presence of the target mRNA, competitive binding between the blocker strand and the mRNA restored Cas9-sgRNA cleavage activity, enabling enzymatic cleavage of adjacent fluorescent dsDNA reporters. Fluorescence measurements were performed at λex/λem = 488/525 nm, yielding a detection limit of 1.40 nmol L⁻¹. This method demonstrated excellent selectivity for Cu/Zn SOD mRNA, as evidenced by its successful application to the detection of Cu/Zn SOD mRNA in real samples.</div></div>\",\"PeriodicalId\":436,\"journal\":{\"name\":\"Talanta Open\",\"volume\":\"12 \",\"pages\":\"Article 100568\"},\"PeriodicalIF\":3.7000,\"publicationDate\":\"2025-09-29\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Talanta Open\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2666831925001699\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"CHEMISTRY, ANALYTICAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Talanta Open","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2666831925001699","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
Sensitive detection Cu/Zn SOD mRNA with CRISPR/Cas9
Cu/Zn SOD mRNA plays a critical role in protecting cells from oxygen toxicity by regulating the expression of the Cu/Zn SOD enzyme. In this work, we developed a biosensor, termed the Cas9-sgRNA/blocker system, for the analysis of Cu/Zn SOD mRNA. The biosensor was assembled by hybridizing a crRNA with a complementary blocker strand, which was then complexed with the Cas9 nuclease. In the presence of the target mRNA, competitive binding between the blocker strand and the mRNA restored Cas9-sgRNA cleavage activity, enabling enzymatic cleavage of adjacent fluorescent dsDNA reporters. Fluorescence measurements were performed at λex/λem = 488/525 nm, yielding a detection limit of 1.40 nmol L⁻¹. This method demonstrated excellent selectivity for Cu/Zn SOD mRNA, as evidenced by its successful application to the detection of Cu/Zn SOD mRNA in real samples.
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