Target-Responsive DNA Hydrogels with Encapsulation and Release Properties Using Programmable CRISPR-Cas12a

Area of Science:

• Biomaterials Science
• Molecular Biology
• Nanotechnology

Background:

• DNA hydrogels offer versatile platforms for drug delivery and sensing.
• Existing hydrogels often lack specificity and rapid response mechanisms.
• CRISPR-Cas gene editing technology provides precise molecular targeting capabilities.

Purpose of the Study:

• To engineer a DNA hydrogel responsive to specific molecular targets.
• To demonstrate programmable, target-induced disassembly and payload release.
• To evaluate the hydrogel's potential in biosensing and therapeutic applications.

Main Methods:

• Assembly of Y-shaped DNA motifs cross-linked with cyanuric acid.
• Assessment of hydrogel integrity using a capillary migration assay.
• Incorporation of CRISPR-Cas12a for target-specific degradation.
• Evaluation of payload release (drug, dye, nanoparticle, protein) upon target detection.
• Testing responsiveness against specific bacterial genomic sequences (Salmonella serotypes).

Main Results:

• The DNA hydrogel demonstrated rapid structural integrity assessment via capillary migration.
• CRISPR-Cas12a enabled selective hydrogel disassembly triggered by specific DNA sequences.
• Controlled release of diverse payloads was achieved, including a drug and a nanoparticle-based agent.
• The hydrogel responded to as few as 50 copies of the target genome (Salmonella typhimurium).
• Differential responsiveness to Salmonella enteritidis and S. typhimurium was observed.

Conclusions:

• CRISPR-responsive DNA hydrogels represent a novel platform for intelligent molecular detection.
• These hydrogels facilitate targeted payload delivery and imaging in response to specific biological triggers.
• The system holds significant potential for advanced biosensing, diagnostics, and therapeutic applications.