A Molecular Translator that Acts by Binding‐Induced DNA Strand Displacement for a Homogeneous Protein Assay

Li, Feng; Zhang, Hongquan; Lai, Chong; Li, Xing‐Fang; Le, X. Chris

doi:10.1002/anie.201202677

Cited by 99 publications

(73 citation statements)

References 35 publications

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“…A broader application of CRISPR technology to non-nucleic acid targets can be achieved by incorporating other molecular translators. 97 CRISPR-Cas systems have been used for DNA and RNA imaging in live cells, 40,41,93 but the target site is limited to repetitive sequences, because of the low copy number of DNA or RNA in one cell and the lack of sufficient signal amplication, 98 CRISPR-Cas-integrated amplication strategies allow for enhanced signal generation at a genomic locus in xed cells and overcome the limitation of CRISPR-Cas probes that could only detect and image repetitive sequences. 42 Amplication techniques using CRISPR-Cas systems have not been demonstrated to function within live cells.…”

Section: Concluding Remarks Outlook and Perspectivesmentioning

confidence: 99%

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CRISPR technology incorporating amplification strategies: molecular assays for nucleic acids, proteins, and small molecules

et al. 2021

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Section: Concluding Remarks Outlook and Perspectivesmentioning

confidence: 99%

“…A broader application of CRISPR technology to non-nucleic acid targets can be achieved by incorporating other molecular translators. 97 …”

Section: Concluding Remarks Outlook and Perspectivesmentioning

confidence: 99%

CRISPR technology incorporating amplification strategies: molecular assays for nucleic acids, proteins, and small molecules

et al. 2021

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“…Many homogenous assays are based on either aptamers [ 6 , 7 ] or DNA-binding proteins [ 8 , 9 ]. Other assays could be quenchbodies [ 10 , 11 ], binding-induced annealing [ 12 , 13 , 14 , 15 , 16 ], or proximity ligation [ 17 , 18 , 19 ]. The latter assays often involve multivalent binding or DNA–protein conjugation.…”

Section: Introductionmentioning

confidence: 99%

A DNA-Based Assay for Digoxin Detection

et al. 2018

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The most common method for quantifying small-molecule drugs in blood samples is by liquid chromatography in combination with mass spectrometry. Few immuno-based assays are available for the detection of small-molecule drugs in blood. Here we report on a homogeneous assay that enables detection of the concentration of digoxin spiked into in a plasma sample. The assay is based on a shift in the equilibrium of a DNA strand displacement competition reaction, and can be performed in 30 min for concentrations above 10 nM. The equilibrium shift occurs upon binding of anti-digoxigenin antibody. As a model, the assay provides a potential alternative to current small-molecule detection methods used for therapeutic drug monitoring.

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“…Remarkably, HRCA offers a detection sensitivity that is four orders of magnitude better than that of regular RCA (10 pM). The PDGF aptamer has a dissociation constant K d of about 0.1 n M 14 and the previously reported structure‐switching fluorescent aptamer biosensor was only able to achieve a detection limit of about 2 n M 17a. Therefore, our integrated strategy offers a dramatically improved detection limit.…”

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confidence: 90%

Biosensing by Tandem Reactions of Structure Switching, Nucleolytic Digestion, and DNA Amplification of a DNA Assembly

Liu

Zhang

et al. 2015

Angew Chem Int Ed

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ϕ29 DNA polymerase (ϕ29DP) is able to carry out repetitive rounds of DNA synthesis using a circular DNA template by rolling circle amplification (RCA). It also has the ability to execute 3'-5' digestion of single-stranded but not double-stranded DNA. A biosensor engineering strategy is presented that takes advantage of these two properties of ϕ29DP coupled with structure-switching DNA aptamers. The design employs a DNA assembly made of a circular DNA template, a DNA aptamer, and a pre-primer. The DNA assembly is unable to undergo RCA in the absence of cognate target owing to the formation of duplex structures. The presence of the target, however, triggers a structure-switching event that causes nucleolytic conversion of the pre-primer by ϕ29DP into a mature primer to facilitate RCA. This method relays target detection by the aptamer to the production of massive DNA amplicons, giving rise to dramatically enhanced detection sensitivity.

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A Molecular Translator that Acts by Binding‐Induced DNA Strand Displacement for a Homogeneous Protein Assay

Cited by 99 publications

References 35 publications

CRISPR technology incorporating amplification strategies: molecular assays for nucleic acids, proteins, and small molecules

CRISPR technology incorporating amplification strategies: molecular assays for nucleic acids, proteins, and small molecules

A DNA-Based Assay for Digoxin Detection

Biosensing by Tandem Reactions of Structure Switching, Nucleolytic Digestion, and DNA Amplification of a DNA Assembly

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