Functionalized Acyclic (l)-Threoninol Nucleic Acid Four-Way Junction with High Stability In Vitro and In Vivo

Area of Science:

• Biomolecular engineering
• Nucleic acid chemistry
• Nanomedicine

Background:

• Oligonucleotides are versatile building blocks for assembling molecules and proteins into defined structures.
• For in vivo applications, oligonucleotide assemblies require high stability, non-toxicity, and non-immunogenicity, which are rarely met by existing derivatives.

Purpose of the Study:

• To investigate acyclic l-threoninol nucleic acid (aTNA) for creating stable, biocompatible oligonucleotide structures for in vivo diagnostics and therapeutics.
• To demonstrate the utility of aTNA-based four-way junctions (4WJs) for facile assembly of functional components.

Main Methods:

• Synthesis and characterization of aTNA four-way junctions (4WJs).
• Assessment of aTNA 4WJ stability in serum.
• Evaluation of cytotoxicity, non-targeted uptake, and innate immune response of aTNA 4WJs.
• Functionalization of aTNA 4WJs with cancer-targeting and serum half-life extension moieties.

Main Results:

• The aTNA 4WJ demonstrated high stability in serum.
• aTNA 4WJs exhibited no non-targeted uptake or cytotoxicity.
• No innate immune response was observed for aTNA 4WJs.
• Functionalized aTNA 4WJs showed potential for targeted delivery and extended circulation in vitro and in vivo.

Conclusions:

• Acyclic l-threoninol nucleic acid (aTNA) is a promising material for constructing stable and biocompatible oligonucleotide assemblies.
• aTNA-based four-way junctions (4WJs) offer a versatile platform for in vivo diagnostics and therapeutics, enabling facile component assembly.
• Functionalized aTNA 4WJs hold potential for targeted cancer treatment and improved pharmacokinetic profiles.