6533b862fe1ef96bd12c6250
RESEARCH PRODUCT
Surface-immobilized DNAzyme-type biocatalysis
Thomas LavergneNicolas SpinelliDavid MonchaudLoic StefanEric Defrancqsubject
StreptavidinSurface PropertiesImmobilized Nucleic AcidsDeoxyribozymeContext (language use)Nanotechnology010402 general chemistryG-quadruplex01 natural sciences[ CHIM ] Chemical Scienceschemistry.chemical_compoundNanobiotechnology[CHIM]Chemical Sciencesheterocyclic compoundsGeneral Materials ScienceComputingMilieux_MISCELLANEOUS010405 organic chemistryDNA Catalytic[CHIM.CATA]Chemical Sciences/Catalysis0104 chemical sciencesG-QuadruplexesPeroxidaseschemistryBiotinylationHelixBiocatalysisOxidation-ReductionDNAdescription
The structure of the double helix of deoxyribonucleic acid (DNA, also called duplex-DNA) was elucidated sixty years ago by Watson, Crick, Wilkins and Franklin. Since then, DNA has continued to hold a fascination for researchers in diverse fields including medicine and nanobiotechnology. Nature has indeed excelled in diversifying the use of DNA: beyond its canonical role of repository of genetic information, DNA could also act as a nanofactory able to perform some complex catalytic tasks in an enzyme-mimicking manner. The catalytic capability of DNA was termed DNAzyme; in this context, a peculiar DNA structure, a quadruple helix also named quadruplex-DNA, has recently garnered considerable interest since its autonomous catalytic proficiency relies on its higher-order folding that makes it suitable to interact efficiently with hemin, a natural cofactor of many enzymes. Quadruplexes have thus been widely studied for their hemoprotein-like properties, chiefly peroxidase-like activity, i.e., their ability to perform hemin-mediated catalytic oxidation reactions. Recent literature is replete with applications of quadruplex-based peroxidase-mimicking DNAzyme systems. Herein, we take a further leap along the road to biochemical applications, assessing the actual efficiency of catalytic quadruplexes for the detection of picomolar levels of surface-bound analytes in an enzyme-linked immunosorbent (ELISA)-type assay. To this end, we exploit an innovative strategy based on the functionalization of DNA by a multitasking platform named RAFT (for regioselectivity addressable functionalized template), whose versatility enables the grafting of DNA whatever its nature (duplex-DNA, quadruplex-DNA, etc.). We demonstrate that the resulting biotinylated RAFT/quadruplex systems indeed acquire catalytic properties that allow for efficient luminescent detection of picomoles of surface-bound streptavidin. We also highlight some of the pitfalls that have to be faced during optimization, notably demonstrating that highly optimized experimental conditions can make DNA pre-catalysts catalytically competent whatever their secondary structures.
year | journal | country | edition | language |
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2014-01-24 |