DNA Condensation Triggered by the Synergistic Self-Assembly of Tetraphenylethylene-Viologen Aggregates and CT-DNA.

Development of small organic chromophores as DNA condensing agents, which explore supramolecular interactions and absorbance or fluorescence-based tracking of condensation and gene delivery processes, is in the initial stages. Herein, we report the synthesis and electrostatic/groove binding interaction-directed synergistic self-assembly of the aggregates of two viologen-functionalized tetraphenylethylene (TPE-V) molecules with CT-DNA and subsequent concentration-dependent DNA condensation process. TPE-V molecules differ in their chemical structure according to the number of viologen units. Photophysical and morphological studies have revealed the interaction of the aggregates of TPE-V in Tris buffer with CT-DNA, which transforms the fibrous network structure of CT-DNA to partially condensed beads-on-a-string-like arrangement with TPE-V aggregates as beads via electrostatic and groove binding interactions. Upon further increasing the concentration of TPE-V, the "beads-on-a-string"-type assembly of TPE-V/CT-DNA complex changes to completely condensed compact structures with 40-50 nm in diameter through the effective charge neutralization process. Enhancement in the melting temperature of CT-DNA, quenching of the fluorescence emission of ethidium bromide/CT-DNA complex, and the formation of induced CD signal in the presence of TPE-V molecules support the observed morphological changes and thereby verify the DNA condensation abilities of TPE-V molecules. Decrease in the hydrodynamic size, increase in the zeta potential value with the addition of TPE-V molecules to CT-DNA, failure of TPE-V/cucurbit(8)uril complex to condense CT-DNA, and the enhanced DNA condensation ability of TPE-V2 with two viologen units compared to TPE-V1 with a single viologen unit confirm the importance of positively charged viologen units in the DNA condensation process. Initial cytotoxicity analysis on A549 cancer and WI-38 normal cells revealed that these DNA condensing agents are non-toxic in nature and hence could be utilized in further cellular delivery studies.

Sequence programmed DNA three-way junctions for templated assembly of fluorescent silver nanoclusters.

Fluorescent silver nanoclusters (AgNCs) templated by DNA are promising label free fluorophores with excellent photostability and tunable optical properties. Most of the reported DNA-nanocluster fluorescent tags comprise of programmed strands for the cluster formation either on the edges as overhangs or as loops on the duplex strands. Herein, we report a design strategy for sequence programmed, DNA three-way junctions (DNA-3WJ), comprising of unhybridized cytosine nucleobases in the 3WJ-center, capable of binding to silver ions and stabilizing the AgNCs. The formation of AgNCs in these DNA-3WJs were confirmed by various spectroscopic and microscopic techniques. 3WJ20-C12 comprising of 12 cytosine bases in the center of the DNA-3WJ, form fluorescent nanoclusters with an emission maximum around 630 nm and 12% fluorescence quantum yield. Control DNA-3WJs with six cytosine bases in the center (3WJ20-C6) and ones without cytosine bases (3WJ20) failed to form fluorescent AgNCs confirming the requirement of central, unhybridized cytosine bases for the stabilization of the nanoclusters. Further, the duplex arms of DNA-3WJs were shown to influence the fluorescent properties of AgNCs by varying the size and stability of the cytosine-loop structure of DNA-3WJs. Metal ion interaction studies shows the selectivity of the 3WJ20-C12/AgNCs towards Hg2+ with sensitivity in the nanomolar range.

Nanosheets and 2D-nanonetworks by mutually assisted self-assembly of fullerene clusters and DNA three-way junctions.

Programmable construction of two dimensional (2D) nanoarchitectures using short DNA strands is of utmost interest in the context of DNA nanotechnology. Previously, we have demonstrated fullerene-cluster assisted self-assembly of short oligonucleotide duplexes into micrometer long, semiconducting nanowires. This report demonstrates the construction of micrometer-sized nanosheets and 2D-nanonetworks from the mutual self-assembly of fullerene nanoclusters with three way junction DNA (3WJ-DNA) and 3WJ-DNA with a 12-mer overhang (3WJ-OH), respectively. The interaction of unique sized fullerene clusters prepared from an aniline appended fullerene derivative, F-An, with two 3WJ-DNAs, namely, 3WJ-20 and 3WJ-30, having 20 and 30 nucleobases, respectively at each strand was characterized using UV-visible absorption, circular dichroism and fluorescence techniques. The morphological characterization of nanosheets embedded with F-An clusters was performed via AFM, TEM and DLS analyses. The programmability and structural tunability of the resultant nanostructures were further demonstrated using 3WJ-OH containing a cytosine rich, single stranded DNA 12-mer overhang, which forms entangled 2D-nanonetwork structures instead of nanosheets due to the differential interaction of F-An nanoclusters with single and duplex strands of 3WJ-OH. Moreover, the selective modification of the cytosine rich sequence present in 3WJ-OH with silver nanoclusters (AgNCs) resulted in significant enhancement in silver nanocluster fluorescence (∼40%) compared to 3WJ-OH/AgNCs owing to the additional stability of AgNCs embedded in 2D nanostructures. This unique strategy of constructing DNA based 2D nanomaterials and their utilization in the integration of functional motifs could find application in the area of DNA nanotechnology and bio-molecular sensing.

Fullerene Cluster Assisted Self-Assembly of Short DNA Strands into Semiconducting Nanowires.

Programmable, hierarchical assembly of DNA nanostructures with precise organisation of functional components have been demonstrated previously with tiled assembly and DNA origami. However, building organised nanostructures with random oligonucleotide strands remains as an elusive problem. Herein, a simple and general strategy, in which nanoclusters of a fullerene derivative act as stapler motifs in bringing ordered nanoscale assembly of short oligonucleotide duplexes into micrometre-sized nanowires, is described. In this approach, the fullerene derivative, by virtue of its amphiphilic structure and unique hydrophobic-hydrophilic balance, pre-assembles to form 3-5 nm sized clusters in a mixture of DMSO-phosphate buffer, which further assists the assembly of DNA strands. The optimum cluster size, availability of DNA anchoring motifs and the nature of the DNA strands controls the structure of these nanomaterials. Furthermore, horizontal conductivity measurements through conductive AFM confirmed the charge transport properties of these nanowires. The current strategy could be employed to organise random DNA duplexes and tiles into functional nanostructures, and hence, open up new avenues in DNA nanotechnology.