Atom‐Modified gDNA Enhances Cleavage Activity of TtAgo Enabling Ultra‐Sensitive Nucleic Acid Testing

Advanced Science, Год журнала: 2024, Номер 11(28)

Опубликована: Май 10, 2024

Abstract The DNA‐guided (gDNA) Argonaute from Thermus thermophilus (TtAgo) has little potential for nucleic acid detection and gene editing due to its poor dsDNA cleavage activity at relatively low temperature. Herein, the of TtAgo is enhanced by using 2′‐fluorine (2′F)‐modified gDNA developes a novel testing strategy. This study finds that with 2′F‐nucleotides 3′‐end (2′F‐gDNA) can promote assembly TtAgo‐guide‐target ternary complex significantly increasing intermolecular force target DNA TtAgo, thereby providing ≈40‐fold enhancement decreasing minimum reaction temperature 65 60 °C. Based on this outstanding advance, strategy proposed, termed FAST, which performed 2′F‐gDNA/TtAgo recognition combining it Bst polymerase amplification. By integrating G‐quadruplex Thioflavin T, FAST assay achieves one‐pot real‐time fluorescence analysis ultra‐sensitivity, limit up 5 copies (20 µL mixture) miR‐21 detection. In summary, an atom‐modification‐based been developed enhancing efficiently, improving practicability establishing TtAgo‐based technology ultra‐sensitivity high‐specificity.

Язык: Английский

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Versatile sensing strategies based on emerging programmable prokaryotic Argonautes: From nucleic acid to non-nucleic acid targets

TrAC Trends in Analytical Chemistry, Год журнала: 2025, Номер unknown, С. 118130 - 118130

Опубликована: Янв. 1, 2025

Язык: Английский

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Advances and applications of G-quadruplex in diagnostics

Microchemical Journal, Год журнала: 2025, Номер unknown, С. 112737 - 112737

Опубликована: Янв. 1, 2025

Язык: Английский

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Comment on: “Atom‐Modified gDNA Enhances Cleavage Activity of TtAgo Enabling Ultrasensitive Nucleic Acid Testing”

Advanced Science, Год журнала: 2025, Номер unknown

Опубликована: Март 8, 2025

The recent article by Zhang et al. piqued the interest. An atom-modification-based strategy is reported to enhance cleavage activity of TtAgo, improving its practicability in TtAgo-based nucleic acid testing. Specifically, 2'-fluorine (2'F)-modified guide DNA (2'F-gDNA) shows significant enhancement TtAgo on double-stranded (dsDNA) increasing melting temperature (Tm) and strengthening binding affinity between 2'F-gDNA targeted dsDNA. These findings are considered important for both molecular diagnostics gene editing. A careful review article, however, raises questions that merit further discussion. After comprehensively reviewing mechanism structure TtAgo-gDNA-target ternary complexes, thoroughly analyzing our results, it believed increased Tm not primary factors activity, speculated 2'F modification at gDNA 3'-end likely influences propagation step. data suggest several details need be addressed improve robustness 2'F-gDNA/TtAgo cleavage.

Язык: Английский

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TtAgo-Coupled-Multiplex-Digtal-RPA-CRISPR/Cas12a (TCMDC) for EGFR Mutations Detection

Talanta, Год журнала: 2024, Номер 283, С. 127162 - 127162

Опубликована: Ноя. 6, 2024

Язык: Английский

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Trends in the use of Argonaute Proteins in Molecular Diagnosis

TrAC Trends in Analytical Chemistry, Год журнала: 2024, Номер unknown, С. 118081 - 118081

Опубликована: Ноя. 1, 2024

Язык: Английский

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Advancements of Prokaryotic Argonautes in Molecular Diagnostics and Further Perspectives

TrAC Trends in Analytical Chemistry, Год журнала: 2024, Номер unknown, С. 118122 - 118122

Опубликована: Дек. 1, 2024

Язык: Английский

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