Highly fluorescent morpholine naphthalimide deoxyuridine nucleotide for the detection of miRNA 24-3P through rolling circle amplification.

Autor: Ravi Kumara GS; Department of Chemistry, Jeonbuk National University, Jeonju 561-756, South Korea. yseo@jbnu.ac.kr., Pandith A; Department of Chemistry, Jeonbuk National University, Jeonju 561-756, South Korea. yseo@jbnu.ac.kr., Seo YJ; Department of Chemistry, Jeonbuk National University, Jeonju 561-756, South Korea. yseo@jbnu.ac.kr.
Jazyk: angličtina
Zdroj: The Analyst [Analyst] 2020 Jul 13; Vol. 145 (14), pp. 4777-4781.
DOI: 10.1039/d0an00723d
Abstrakt: In this study we synthesized the nucleotide dUrkTP, a highly fluorescent naphthalimide deoxyuridine triphosphate that undergoes aggregation-induced emission (AIE). We incorporated and extended dUrkTP during the primer extension of DNA mediated by DNA polymerase, and also in the rolling circle amplification of DNA mediated by Phi29 polymerase. Accordingly, we could use this fluorescent nucleotide for the detection of microRNA 24-3P, a biomarker of porcine reproductive and respiratory syndrome virus. The direct labeling system obtained during rolling circle DNA amplification exhibited increased fluorescence, due to AIE of the dUrkTP residue upon gel formation, thereby allowing the detection of miRNA 24-3P. This direct labeling system facilitated the simple and inexpensive detection of miRNA 24-3P with high sensitivity (limit of detection: 3.58 fM) and selectivity.
Databáze: MEDLINE