Engineering Micrometer-Sized DNA Tracks for High-Speed DNA Synthesis and Biosensing.
Angew Chem Int Ed Engl
; 59(51): 22947-22951, 2020 12 14.
Article
em En
| MEDLINE
| ID: mdl-33007137
ABSTRACT
φ29 DNA polymerase (Polφ29) is capable of synthesizing long-chain single-stranded (ss) DNA molecules by copying the sequence of a small ss circular DNA template (ssCDT) in a process known as rolling circle amplification (RCA). The use of a ssCDT in RCA, however, comes with a key drawback the rate of DNA synthesis is significantly reduced. We hypothesize that this issue can be overcome using a very long linear ssDNA template with a repeating sequence. To test this idea, we engineered a DNA assembly, which we denote "micrometer-sized DNA track" (µDT). This µDT, with an average length of ≈13.5â
µm, is made of a long chain DNA with a primer-binding domain at its 3' end and ≈1000 repeating sequence units at its 5' end, each carrying a DNA anchor. We find that Polφ29 copies µDT at a speed ≈5-time faster than it does a related ssCDT. We use this to design a simple all-in-one printed paper device for rapid and sensitive detection of microRNA let-7. This paper sensor is capable of detecting 1â
pM let-7a in 10â
minutes.
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Coleções:
01-internacional
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MEDLINE
Assunto principal:
DNA
/
Técnicas Biossensoriais
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Técnicas de Amplificação de Ácido Nucleico
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DNA Polimerase Dirigida por DNA
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article