Guided magnetic nanostructures for targeted and high-throughput intracellular delivery
Inventors
Weiss, Paul S. • Tseng, Hsian-rong • XU, Xiaobin • Wattanatorn, Natcha • Yang, Qing • Jonas, Steven J.
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Assignees
University of California San Diego UCSD
University of California, San Diego (UCSD)The University of California, San Diego (UCSD) is a leading public research university located in La Jolla, California. Known for its innovative and interdisciplinary approach, UCSD offers a wide range of undergraduate, graduate, and professional programs across various fields. The university is committed to fostering a diverse and inclusive community, promoting sustainability, and driving social mobility through education, research, and public service. UCSD is recognized for its contributions to research and innovation, particularly in areas such as climate science, health innovation, and artificial intelligence.
The University of California, San Diego (UCSD) is a leading public research university located in La Jolla, California. Known for its innovative and interdisciplinary approach, UCSD offers a wide range of undergraduate, graduate, and professional programs across various fields. The university is committed to fostering a diverse and inclusive community, promoting sustainability, and driving social mobility through education, research, and public service. UCSD is recognized for its contributions to research and innovation, particularly in areas such as climate science, health innovation, and artificial intelligence.
Publication Number
US-12359220-B2
Publication Date
2025-07-15
Expiration Date
2039-01-29
Abstract
A method of transporting biomolecular cargo intracellularly into cells includes the operations of providing magnetic nanostructures (e.g., nanospears, nanostars, nanorods, and other nanometer-sized structures) carrying the biomolecular cargo thereon and applying an external magnetic field to move the magnetic nanostructures into physical contact with at least some of the cells (or the cells into the magnetic nanostructures). The magnetic nanostructures move into physical contact with a single cell, a subset of cells, or all cells. The external magnetic field may be applied by a moving permanent magnet although an electromagnetic may also be used. The biomolecular cargo may include a molecule, a plurality of molecules, or higher order biological constructs. For example, the biological construct may include DNA plasmids, small interfering RNA, proteins, or targeted nuclease gene-editing cargo such as zinc-finger nucleases, transcription activator-like effector nucleases, Cas9 protein, Cas9 mRNA, and associated guide RNA sequences.
Core Innovation
The invention relates to the design and fabrication of magnetic nanostructures that can be readily configured for single-cell modification and progressively scaled for direct and highly efficient intracellular delivery of biomolecular cargo. These biocompatible nanomaterials are guided precisely to target cells by manipulation of locally applied external magnetic fields without the need for chemical propellants. The magnetic nanostructures may include various shapes such as nanospears, nanostars, nanorods, and other nanometer-sized structures, which have at least one sharp end, tip, or surface to facilitate penetration into cellular membranes or walls.
The method includes providing magnetic nanostructures carrying biomolecular cargo and applying an external magnetic field to move the magnetic nanostructures into physical contact with single cells, subsets of cells, or all cells. The targeting of particular cells or groups of cells is achieved by positioning and/or manipulation of the external magnetic field. Biomolecular cargo may include DNA plasmids, proteins, small interfering RNA, or targeted nuclease gene-editing cargo such as zinc-finger nucleases, TALENs, Cas9 protein, Cas9 mRNA, and guide RNA sequences. The nanostructures are biocompatible and may be biodegradable by the target cells over time.
The problem being solved is the challenge of achieving high-throughput and targeted intracellular delivery of biomolecules that is safe, efficient, cost-effective, and minimally stressful to cells. Existing approaches such as viral vectors, electroporation, chemical reagents, and other membrane-disruption-based methods are often costly, toxic, or inefficient. Current nano-needle platforms involve substrates that make cell release difficult, and catalytic propulsion-based nano/micromotor systems have limited guidance precision and biocompatibility. The invention addresses these issues by providing magnetic nanostructures that can be magnetically guided without chemical propellants, allowing precise, scalable, and controlled intracellular delivery of various biomolecules.
Claims Coverage
The patent includes one independent claim describing a method of transporting biomolecular cargo intracellularly into cells in a microfluidic channel using magnetic nanostructures. The main inventive features cover the aspects of the magnetic nanostructures, their orientation, positioning, and interaction with cells under a controlled magnetic field.
Method of intracellular biomolecular cargo transport using magnetic nanostructures in a microfluidic channel
A method comprising providing a microfluidic channel with a plurality of magnetic nanostructures carrying biomolecular cargo thereon, wherein the nanostructures possess at least one pointed end or tip; flowing a fluid containing the magnetic nanostructures within the microfluidic channel; applying an external magnetic field to maintain the nanostructures in a substantially stationary state along the channel width and orient them so that the pointed ends face opposite the flow direction; and flowing cells within the microfluidic channel to physically contact the pointed ends of the magnetic nanostructures.
The inventive features focus on the use of magnetically guided nanostructures with pointed tips immobilized and oriented in microfluidic flow channels to contact cells and deliver biomolecular cargo. This allows effective and targeted intracellular delivery in a controlled microenvironment.
Stated Advantages
The magnetic nanostructures can be guided precisely to target cells without chemical propellants, avoiding potential toxicities associated with catalytic propulsion.
The approach enables high-throughput and targeted intracellular delivery with high efficiency and minimal impact on cell viability and metabolism.
The magnetic nanostructures are biocompatible and biodegradable by the target cells over time.
The method is scalable for large populations of cells and compatible with good manufacturing practices, enabling batch processing for ex vivo gene-modified cell therapies.
The magnetic nanostructures provide precise control over position, orientation, and speed, enabling targeted single-cell or subpopulation intracellular delivery.
Documented Applications
Targeted intracellular delivery of biomolecular cargo such as plasmid DNA, RNA, proteins, and gene-editing agents into single cells or subsets of cells in vitro and ex vivo.
High-throughput transfection of large populations of cells in laboratory culture environments using volleys of magnetic nanostructures.
Targeted transfection of specific cell populations located at distinct geographic regions on a substrate, enabling localized delivery of different genetic payloads.
Potential application in molecular cell biology studies and translational medicine, including gene therapy and cell and tissue engineering.
Use within microfluidic channels to deliver biomolecular cargo by maintaining magnetic nanostructures stationary and oriented to interact with flowing cells.
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