by A group of researchers from various fields of engineering and physics has developed a novel method called CRISPR-powered optothermal nanotweezers (CRONT) that combines the power of optothermal manipulation with Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technology. This innovative approach enhances the detection and manipulation of nanoparticles, particularly in the field of bionanotechnology.
Traditional optical tweezers, which use light momentum transformation, have limitations in nanoparticle regulation. Optothermal nanotweezers, on the other hand, employ optical-induced thermodynamic forces to precisely manipulate nanoparticles at the micron-scale, with sub-micron precision. Compared to traditional optical tweezers, optothermal tweezers require lower power density, making them more suitable for biological detection and minimizing adverse effects on biological samples.
In this study, the researchers successfully enriched DNA functionalized gold nanoparticles, CRISPR-associated proteins, and DNA strands by harnessing diffusiophoresis and thermo-osmotic flows for optothermal excitation. They also enhanced the single-nucleotide polymorphism detection capabilities of CRISPR-associated proteins, introducing a new method to observe nucleotide cleavage. This methodology has universal applications in point-of-care diagnostics, biophotonics, and bionanotechnology.
To enable CRONT, the researchers designed a microfluidic chamber with a thin layer of gold film. When the gold film was irradiated with laser illumination, it generated a temperature field that surrounded the laser spot. The optimal conditions for CRISPR reactions were determined, and cleavage of the DNA-gold nanofilm conjugate was initiated using dark-field microscopy. The researchers also incorporated a nonionic polymer, polyethylene glycol (PEG), as a biological surfactant to enhance biocompatibility.
The presence of multiple nanoparticles with varying thermophoretic mobility led to the generation of a distinct solute concentration, resulting in an interaction known as the diffusiophoretic force. This systematic investigation demonstrated the potential of CRONT for biomolecular identification.
Fluorescence labeling was used to study the aggregation behaviors of proteins and DNA. It was observed that the accumulation rate of single-stranded DNA was higher than that of double-stranded DNA, while protein accumulations showed a tendency to form slight ring-like structures. The accumulation rate increased with higher laser power.
The CRONT system successfully identified DNAs at the single molecule level for single nucleotide polymorphisms with high sensitivity and specificity. By incorporating diffusiophoresis and thermo-osmotic flows into the boundary layer of an optothermal responsive film, the researchers demonstrated the precise regulation of CRONT at the nanoscale.
CRONT has immense potential in advancing the understanding of complex biological processes and can be used as a versatile detection probe in biomedical research, drug discovery, and disease diagnostics. This innovative methodology opens up new possibilities for efficient nanoparticle manipulation and paves the way for further advancements in bionanotechnology.
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1. Source: Coherent Market Insights, Public sources, Desk research
2. We have leveraged AI tools to mine information and compile it
Money Singh is a seasoned content writer with over four years of experience in the market research sector. Her expertise spans various industries, including food and beverages, biotechnology, chemicals and materials, defense and aerospace, consumer goods, etc.
