Direct incorporation and extension of a fluorescent nucleotide through rolling circle DNA amplification for the detection of microRNA 24-3P

• Published in: Bioorganic & medicinal chemistry letters Vol. 28; no. 11; pp. 2035 - 2038
• Main Authors: Le, Binh Huy, Seo, Young Jun
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 15.06.2018
• Subjects: Anthracenes - chemical synthesis; Anthracenes - chemistry; Direct labeling; DNA, Circular - genetics; Dose-Response Relationship, Drug; Fluorescence; Fluorescent Dyes - chemical synthesis; Fluorescent Dyes - chemistry; Fluorescent nucleotide; Humans; MicroRNAs - analysis; miRNA; Molecular Structure; Nucleic Acid Amplification Techniques; Pyrenes - chemical synthesis; Pyrenes - chemistry; RCA; Structure-Activity Relationship; Thiophenes - chemical synthesis; Thiophenes - chemistry; Virus; Virus; Direct labeling; RCA; miRNA; Fluorescent nucleotide
• ISSN: 0960-894X; 1464-3405
• DOI: 10.1016/j.bmcl.2018.04.058

[Display omitted] •The thiophene-based fluorescent nucleotide (dUthioTP) could highly incorporate and extended into the rolling circle DNA product.•dUthioTP fluorescent nucleotide could be used for the detection of miRNA 24-3P which is related PRRSV.•dUthioTP-RCA direct labeling system is a very simple and cost-efficient method for the detection miRNA 24-3P. We designed and synthesized several fluorescent nucleotides from thiophene, anthracene and pyrene, which have different sizes, and screened their incorporation and extension capability during the rolling circle amplification of DNA. The thiophene-based fluorescent nucleotide (dUthioTP) could highly incorporate and extended into the rolling circle DNA product, while other fluorescent nucleotides (dUanthTP, and dUpyrTP) could not. This dUthioTP fluorescent nucleotide could be used for the detection of miRNA 24-3P, which is related PRRSV. This direct labeling system during rolling circle DNA amplification exhibited an increased fluorescence signal showing gel formation for the detection of miRNA 24-3P. This direct labeling system is a very simple and cost-efficient method for the detection miRNA 24-3P and also exhibited highly sensitive and selective detection properties.