Preparation of Stable Branched DNA Nanostructures: Process of Cooperative Self-Assembly

The construction of functionalizable branched DNA (bDNA) relies on the designing of oligonucleotides and exploitation of their complementary chemistries. The stability of these structures largely depends on the hybridization specificity of the contributing oligonucleotides. However, most of the bDNA structures are not found suitable for in vivo application due to poor yield owing to uncharacterized hybridization efficiency and instability in biological fluids. In this report, our group has explored a mechanistic way for studying the hybridization pathway of genomic sequence derived oligonucleotides that are self-assembled to fabricate robust bDNA structures. The effect of change in nucleotide sequences on bDNA stability was studied by taking oligonucleotides derived from primers of different genes. Additionally, the stability of the bDNA in solutions with different pH, salts, and DNaseI which mimics physiological environment was reported. It was found that genomic sequence derived oligonucleotides self-assembled in a cooperative manner to yield the designed bDNAs, which are stable in physiological environment.