Circular RNA oligonucleotides: enzymatic synthesis and scaffolding for nanoconstruction

Area of Science:

• Biochemistry
• Molecular Biology
• Nanotechnology

Background:

• Circular RNAs (circRNAs) are gaining attention for their diverse biological roles.
• Efficient synthesis of short, defined circRNAs remains a challenge.
• Nucleic acid nanotechnology utilizes self-assembling structures for various applications.

Purpose of the Study:

• To develop an efficient method for synthesizing monomeric circular RNAs (circRNAs) between 16-44 nucleotides.
• To utilize synthesized circRNAs as scaffolds for constructing novel nucleic acid nanostructures.

Main Methods:

• A novel DNA dumbbell splinting strategy combined with T4 DNA ligation was employed for circRNA synthesis.
• The synthesized 44 nt circRNA was used as a scaffold for building hybrid RNA:DNA and pure RNA:RNA double crossover tiles.
• Assembly of these tiles into nucleic acid nanotubes and flat arrays was achieved.

Main Results:

• Efficient synthesis of monomeric circRNAs in the 16-44 nt size range was achieved.
• Successful construction of hybrid RNA:DNA and pure RNA:RNA double crossover tiles using circRNA scaffolds.
• Formation of self-assembled nucleic acid nanotubes and flat arrays from these tiles.

Conclusions:

• The novel DNA dumbbell splinting strategy provides an efficient route to synthesize short monomeric circRNAs.
• Synthesized circRNAs can serve as versatile scaffolds for constructing complex nucleic acid nanostructures.
• This work expands the toolkit for nucleic acid nanotechnology and circRNA research.