New Fluorescent Nanoparticles for Ultrasensitive Detection of Nucleic Acids by Optical Methods

Mulberg, Mads Westergaard; Taskova, Maria; Thomsen, Rasmus P.; Okholm, Anders Hauge; Kjems, Jørgen; Astakhova, Kira

doi:10.1002/cbic.201700125

Cited by 3 publications

(2 citation statements)

References 32 publications

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“…These results showed that the design can be tuned to enhance the sensor stability without compromising the sensitivity of the sensor. Given that the vast majority of sensing approaches available today require either amplification of target (enzymatic or nonenzymatic), labeling of target, or some sort of signal amplification such as the use of nanomaterials or nanocomposites to reach low nM to fM detection limits, − the sensing approach that we demonstrated here offers an ultrasensitive detection of nucleic acids in a simpler format.…”

Section: Resultsmentioning

confidence: 99%

Single-Molecule FRET-Based Dynamic DNA Sensor

2021

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Selective and sensitive detection of nucleic acid biomarkers is of great significance in early-stage diagnosis and targeted therapy. Therefore, the development of diagnostic methods capable of detecting diseases at the molecular level in biological fluids is vital to the emerging revolution in the early diagnosis of diseases. However, the vast majority of the currently available ultrasensitive detection strategies involve either target/signal amplification or involve complex designs. Here, using a p53 tumor suppressor gene whose mutation has been implicated in more than 50% of human cancers, we show a background-free ultrasensitive detection of this gene on a simple platform. The sensor exhibits a relatively static mid-FRET state in the absence of a target that can be attributed to the time-averaged fluorescence intensity of fast transitions among multiple states, but it undergoes continuous dynamic switching between a low-and a high-FRET state in the presence of a target, allowing a high-confidence detection. In addition to its simple design, the sensor has a detection limit down to low femtomolar (fM) concentration without the need for target amplification. We also show that this sensor is highly effective in discriminating against single-nucleotide polymorphisms (SNPs). Given the generic hybridization-based detection platform, the sensing strategy developed here can be used to detect a wide range of nucleic acid sequences enabling early diagnosis of diseases and screening genetic disorders.

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Section: Resultsmentioning

confidence: 99%

Single-Molecule FRET-Based Dynamic DNA Sensor

2021

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“…However, chromatography is not suitable for biosensing and needs complex equipment; a needle microelectrode needs to be inserted into the tumor site in the electrochemistry method, which is invasive, and furthermore, this technique cannot be used for cellular sensing. The luminescence method is noninvasive, sensitive, highly selective, and convenient and can be employed in vitro and in vivo to map the heterogeneous distribution of oxygen …”

Section: Methodsmentioning

confidence: 91%

Compact Conjugated Polymer Dots with Covalently Incorporated Metalloporphyrins for Hypoxia Bioimaging

Fang

Liu

et al. 2018

ChemBioChem

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Hypoxia is closely related to multiple diseases, especially in tumors, which increases the aggressiveness and drug resistance of cancer cells. Precise hypoxia imaging is of great significance for cancer diagnosis and the evaluation of therapeutic effects. A kind of hydrophobic polymer (i.e., PFPtTFPP) as an imaging probe for hypoxia with fluorene as an energy donor and an oxygen‐sensitive PtII porphyrin as an energy acceptor was developed. Compact polymer dots (Pdots) with a small size were prepared by nanoprecipitation. The PFPtTFPP Pdots showed excellent hypoxia sensing in solution with high sensitivity and full reversibility. The emission intensity, quantum yields, lifetime, and single‐particle brightness significantly increased under hypoxia conditions. Remarkably, hypoxia imaging in vitro and in vivo was realized, and a clear increase in brightness was observed under hypoxia conditions and in the tumor area. Excellent hypoxia imaging ability is beneficial to potential applications in cancer diagnosis.

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Complementary Oligonucleotide Conjugated Multicolor Carbon Dots for Intracellular Recognition of Biological Events

Srivastava

Misra

Bangru

et al. 2020

ACS Appl. Mater. Interfaces

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By using complementary DNA sequences as surface ligands, we selectively allow two individual diffusing “dual-color” carbon dots to interact in situ and in vitro. Spontaneous nanoscale oxidation of surface-abundant nitroso-/nitro-functionalities leads to two distinctly colored carbon dots (CD) which are isolated by polarity driven chromatographic separation. Green- and red-emitting carbon dots (gCD and rCD) were decorated by complementary single-stranded DNAs which produce a marked increase in the fluorescence emission of the respective carbon dots. Mutual colloidal interactions are achieved through hybridization of complementary DNA base pairs attached to the respective particles, resulting in quenching of their photoluminescence. The observed post-hybridization quenching is presumably due to a combined effect from an aggregation of CDs post duplex DNA formation and close proximity of multicolored CDs, having overlapped spectral regions leading to a nonradiative energy transfer process possibly released as heat. This strategy may contribute to the rational design of mutually interacting carbon dots for a better control over the resulting assembly structure for studying different biological phenomenon including molecular cytogenetics. One of the newly synthesized CDs was successfully used to image intracellular location of GAPDH mRNA using an event of change in fluorescence intensity (FI) of CDs. This selectivity was introduced by conjugating an oligonucleotide harboring complementary sequence to GAPDH mRNA. FI of this conjugated carbon dot, rCD-GAPDH, was also found to decrease in the presence of Ca2+, varied in relation to H+ concentrations, and could serve as a tool to quantify the intracellular concentrations of Ca2+ and pH value (H+) which can give important information about cell survival. Therefore, CD-oligonucleotide conjugates could serve as efficient probes for cellular events and interventions.

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New Fluorescent Nanoparticles for Ultrasensitive Detection of Nucleic Acids by Optical Methods

Cited by 3 publications

References 32 publications

Single-Molecule FRET-Based Dynamic DNA Sensor

Single-Molecule FRET-Based Dynamic DNA Sensor

Compact Conjugated Polymer Dots with Covalently Incorporated Metalloporphyrins for Hypoxia Bioimaging

Complementary Oligonucleotide Conjugated Multicolor Carbon Dots for Intracellular Recognition of Biological Events

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