Tumor‐Targeting Cholesterol‐Decorated DNA Nanoflowers for Intracellular Ratiometric Aptasensing

• Published in: Advanced materials (Weinheim) 2021-03, Vol.33 (11), p.e2007738-n/a
• Main Authors: Kim, Nayoung, Kim, Eunjung, Kim, Hyemin, Thomas, Michael R., Najer, Adrian, Stevens, Molly M.
• Format: Article
• Language: English
• Subjects: Abundance; Adenosine triphosphate; Adenosine Triphosphate - analysis; Aptamers, Nucleotide - chemistry; aptasensors; Biomimetics; Biomolecules; Biosensing Techniques - methods; Cell Line, Tumor; Cholesterol; Cholesterol - analysis; Cholesterol - chemistry; Crystallization; Deoxyribonucleic acid; DNA; DNA - chemistry; DNA flowers; Energy transfer; Fluorescence Resonance Energy Transfer - methods; Förster resonance energy transfer; Humans; Materials science; Nanocomposites; Nanostructures - chemistry; Prostatic Neoplasms - diagnosis; ratiometric aptasensing; rolling circle amplification; Synergistic effect; Target detection; Tumors
• ISSN: 0935-9648; 1521-4095; 1521-4095
• DOI: 10.1002/adma.202007738

Probing endogenous molecular profiles is of fundamental importance to understand cellular function and processes. Despite the promise of programmable nucleic‐acid‐based aptasensors across the breadth of biomolecular detection, target‐responsive aptasensors enabling intracellular detection are as of yet infrequently realized. Several challenges remain, including the difficulties in quantification/normalization of quencher‐based intensiometric signals, stability issues of the probe architecture, and complex sensor operations often necessitating extensive structural modeling. Here, the biomimetic crystallization‐empowered self‐assembly of a tumor‐targetable DNA–inorganic hybrid nanocomposite aptasensor is presented, which enables Förster resonance energy transfer (FRET)‐based quantitative interpretation of changes in the cellular target abundance. Leveraging the design programmability and high‐throughput fabrication of rolling circle amplification‐driven DNA nanoarchitecture, this designer platform offers a method to self‐assemble a robust nanosensor from a multifunctionality‐encoded template that includes a cell‐targeting aptamer, a ratiometric aptasensor, and a cholesterol‐decorating element. Taking prostate cancer cells and intracellular adenosine triphosphate molecules as a model system, a synergistic effect in the targeted delivery by cholesterol and aptamers, and the feasibility of quantitative intracellular aptasensing are demonstrated. It is envisioned that this approach provides a highly generalizable strategy across wide‐ranging target systems toward a biologically deliverable nanosensor that enables quantitative monitoring of the abundance of endogenous biomolecules. A programmable DNA–inorganic hybrid nanocomposite (DNA nanoflower) intracellular aptasensor with sequence‐encoded multifunctional features is realized by implementing enzymatic amplification‐driven biomimetic crystallization. This platform is capable of ratiometric and quantitative intracellular detection upon targeted delivery and holds substantial promise as strategically self‐assembled DNA nanosensors for interrogating cellular analytes whose changes in abundance are explained as a continuous function.