ABSTRACT
G-rich telomeric DNA plays a major role in the stabilization of chromosomes and can fold into a plethora of different G-quadruplex structures in the presence of mono- and divalent cations. The reversed human telomeric DNA sequence (5'-(GGG ATT)4; RevHumTel) was previously shown to have interesting properties that can be exploited for chemical sensing and as a chemical switch in DNA nanotechnology. Here, we analyze the specific G-quadruplex structures formed by RevHumTel in the presence of K+, Na+, Mg2+ and Ca2+ cations using circular dichroism spectroscopy (CDS) and Förster resonance energy transfer (FRET) based on fluorescence lifetimes. CDS is able to reveal strand and loop orientations, whereas FRET gives information about the distances between the 5'-end and the 3'-end, and also, the number of G-quadruplex species formed. Based on this combined information we derived specific G-quadruplex structures formed from RevHumTel, i.e., a chair-type and a hybrid-type G-quadruplex structure formed in presence of K+, whereas Na+ induces the formation of up to three different G-quadruplexes (a basket-type, a propeller-type and a hybrid-type structure). In the presence of Mg2+ and Ca2+ two different parallel G-quadruplexes are formed (one of which is a propeller-type structure). This study will support the fundamental understanding of the G-quadruplex formation in different environments and a rational design of G-quadruplex-based applications in sensing and nanotechnology.
Subject(s)
G-Quadruplexes , Humans , Salts , Base Sequence , Cations, Divalent , Potassium/chemistry , Circular Dichroism , Telomere/genetics , DNA/chemistry , Sodium/chemistryABSTRACT
Rapid and sensitive detection of biomarkers enables monitoring patients' health status and can enhance the early diagnosis of deadly diseases. In this work, we have developed a new colorimetric platform based on spherical nucleic acid (SNA) and G-quadruplex DNAzymes for the identification of specific miRNAs. The simple hybridization between the target miRNA and two capture probes (capture probe 1 located at AuNP surface and free capture probe 2) is the working principle of this biosensor. The hybridization and duplex formation among probes and miRNAs led to a significant decrease in the intensity of color change. A linear relationship between the decrease of colorimetric signal and the amount of target molecules was witnessed from 1 to 100 nM for miRNA-155. Using this method, we were able to detect concentrations of miRNA-155 as low as 0.7 nM. Furthermore, the proposed sensing platform can be utilized profitably to detect miRNA-155 in real human serum samples. We further investigated the applicability of the proposed method in a microfluidic system which displayed promising results. In this project, A G-quadruplex based SNAzyme was constructed to provide a fast and simple colorimetric method for miRNA detection. The SNAzyme actually employed as both target recognition element and catalytic nano labels for colorimetric detection.
Subject(s)
Biosensing Techniques , DNA, Catalytic , G-Quadruplexes , MicroRNAs , Biosensing Techniques/methods , Colorimetry/methods , DNA, Catalytic/genetics , HumansABSTRACT
Biochemical properties and flexibility of nitrogenous bases allow DNA to fold into higher-order structures. Among different DNA secondary structure, G-quadruplexes (tetrapelexes-G4) - which are formed in guanine rich sequences - have gained more attention because of their biological significance, therapeutic intervention, and application in molecular device and biosensor. G4-quadruplex studies categorize into three main fields, in vivo, in vitro, and in silico. The in vitro field includes G4 synthetic oligonucleotides. This review focuses on the G-quadruplex synthetic aptamers structure features and considers the applicability of G4-aptamers for cancer biomarkers detection. Various biosensing methods will be reviewed based on G-quadruplex aptamers for cancer detection.
