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Signal Amplification Technologies for the Detection of Nucleic Acids: from Cell-Free Analysis to Live-Cell Imaging

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Abstract

Due to their unique properties, such as programmability, ligand-binding capability, and flexibility, nucleic acids can serve as analytes and/or recognition elements for biosensing. To improve the sensitivity of nucleic acid-based biosensing and hence the detection of a few copies of target molecule, different modern amplification methodologies, namely target-and-signal-based amplification strategies, have already been developed. These recent signal amplification technologies, which are capable of amplifying the signal intensity without changing the targets’ copy number, have resulted in fast, reliable, and sensitive methods for nucleic acid detection. Working in cell-free settings, researchers have been able to optimize a variety of complex and quantitative methods suitable for deploying in live-cell conditions. In this study, a comprehensive review of the signal amplification technologies for the detection of nucleic acids is provided. We classify the signal amplification methodologies into enzymatic and non-enzymatic strategies with a primary focus on the methods that enable us to shift away from in vitro detecting to in vivo imaging. Finally, the future challenges and limitations of detection for cellular conditions are discussed.

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Abbreviations

ABTS:

2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonicacid

AP:

Alkaline phosphatase

AuNPs:

Gold nanoparticles

bDNA:

Branched DNA

CHA:

Catalytic hairpin assembly

DNA:

Deoxyribonucleic acid

DSN:

Duplex-specific nuclease

dsDNA:

Double-stranded DNA

EDC:

Entropy-driven catalysis

FISH:

Fluorescence in situ hybridization

FRET:

Fluorescence resonance energy transfer

GO:

Graphene oxide

HCR:

Hybridization chain reaction

ISH:

In situ hybridization

LOD:

Limit of detection

LSPR:

Localized surface plasmon resonance

MB:

Molecular beacon

MMPs:

Magnetic microparticles

NC:

Nanocluster

PCR:

Polymerase chain reaction

PMP:

Polymer microparticles

PO:

Horseradish peroxidase

QDs:

Quantum dots

SERS:

Surface-enhanced Raman scattering

SNPs:

Single nucleotide polymorphisms

TSA:

Tyramid signal amplification

TSDRs:

Toehold-mediated DNA strand displacement reactions

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Acknowledgments

The authors acknowledge the financial support of this investigation by the Research Council of Shahid Bahonar University of Kerman (Kerman, Iran).

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  1. Department of Biology, Faculty of Science, Shahid Bahonar University of Kerman, Kerman, Iran

    Tahereh Fozooni, Hadi Ravan & Hosseinali Sasan

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  1. Tahereh Fozooni
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  2. Hadi Ravan
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  3. Hosseinali Sasan
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Correspondence to Hadi Ravan.

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Fozooni, T., Ravan, H. & Sasan, H. Signal Amplification Technologies for the Detection of Nucleic Acids: from Cell-Free Analysis to Live-Cell Imaging. Appl Biochem Biotechnol 183, 1224–1253 (2017). https://doi.org/10.1007/s12010-017-2494-4

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