Arrest of Rolling Circle Amplification by Protein‐Binding DNA Aptamers |
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| Authors: | Lida Wang Kha Tram Dr Monsur M Ali Prof Bruno J Salena Prof Jinghong Li Prof?Dr Yingfu Li |
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| Affiliation: | 1. Department of Biochemistry and Biomedical Sciences and Department of Chemistry and Chemical Biology, McMaster University, 1280 Main St. W., Hamilton, ON, L8S 4K1 (Canada);2. Department of Chemistry and Beijing Key Laboratory for Microanalytical Methods and Instrumentation, Tsinghua University, Beijing, 100084 (P. R. China);3. Division of Gastroenterology, Department of Medicine, McMaster University, 1280 Main St. W. Hamilton, ON, L8S 4K1 (Canada) |
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| Abstract: | Certain DNA polymerases, such as ?29 DNA polymerase, can isothermally copy the sequence of a circular template round by round in a process known as rolling circle amplification (RCA), which results in super‐long single‐stranded (ss) DNA molecules made of tandem repeats. The power of RCA reflects the high processivity and the strand‐displacement ability of these polymerases. In this work, the ability of ?29DNAP to carry out RCA over circular templates containing a protein‐binding DNA aptamer sequence was investigated. It was found that protein–aptamer interactions can prevent this DNA polymerase from reading through the aptameric domain. This finding indicates that protein‐binding DNA aptamers can form highly stable complexes with their targets in solution. This novel observation was exploited by translating RCA arrest into a simple and convenient colorimetric assay for the detection of specific protein targets, which continues to showcase the versatility of aptamers as molecular recognition elements for biosensing applications. |
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| Keywords: | aptamers biosensors molecular competition protein detection rolling circle amplification |
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