Vector-free intracellular delivery by reversible permeabilisation
Inventors
Maguire, Michael • O'Dea, Shirley
Assignees
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Publication Number
US-12060570-B2
Publication Date
2024-08-13
Expiration Date
Abstract
The invention provides a solution to the problem of transfecting non-adherent cells. Devices and delivery compositions containing ethanol and an isotonic salt solution are used for delivery of compounds and compositions to non-adherent cells.
Core Innovation
The invention relates to a system and an in vitro method for vector-free intracellular delivery by contacting a population of cells with a volume of aqueous delivery solution that is isotonic and contains a payload comprising a gene editing composition. The method forms a spray by propelling the aqueous solution with gas and contacts the cells with an atomized delivery solution delivered by a delivery solution applicator.
The system includes a housing configured to receive a plate with a well containing a population of cells, and a differential pressure applicator configured to apply a differential pressure to the well. A stop solution applicator delivers a stop solution to the well, and a culture medium applicator delivers culture medium to the well, supporting controlled delivery and recovery following contact with the atomized delivery solution.
The delivery solution further includes an alcohol at greater than 5 percent (v/v) together with the isotonic aqueous phase and the gene editing composition payload. The differential pressure applicator and gas-propelled atomization are combined with downstream culture-medium delivery, enabling contact of cells with gene-editing payloads delivered as an atomized spray while using a stop solution and culture medium to manage the delivery environment.
Claims Coverage
The independent claims are clm-00001 and clm-00028. Across these claims, the coverage centers on a cell-culture system and a corresponding in vitro method that use differential pressure and atomized isotonic aqueous delivery propelled by gas, with a gene-editing payload in an alcohol-containing delivery solution, followed by stop-solution and culture-medium delivery.
Differential pressure cell-contact system with atomized isotonic gene-editing delivery
A system comprising a plate with a well containing a population of cells in a housing; a differential pressure applicator configured to apply a differential pressure to the well; a delivery solution applicator configured to deliver atomized delivery solution to the well; and a reservoir fluidically coupled to the delivery solution applicator, the reservoir including a delivery solution having an isotonic aqueous solution with a payload comprising a gene editing composition, where the aqueous solution is contacted with the cells by gas propelling the aqueous solution to form a spray.
Stop solution and culture medium delivery after atomized contact
The system further includes a stop solution applicator configured to deliver a stop solution to the well and a culture medium applicator configured to deliver a culture medium to the well, in the context of contacting cells with the atomized delivery solution containing the gene editing composition.
Alcohol-containing isotonic atomized gene-editing in vitro method propelled by gas
An in vitro method providing a population of cells within a well of a system including a housing; a differential pressure applicator configured to apply a differential pressure to the well; a delivery solution applicator configured to deliver atomized delivery solution to the well; and a reservoir fluidically coupled to the delivery solution applicator, the reservoir including a delivery solution with an aqueous solution isotonic to include a payload comprising a gene editing composition and an alcohol at greater than 5 percent (v/v), and contacting the population of cells with a volume of aqueous solution by gas propelling the aqueous solution to form a spray.
The claim coverage is directed to contacting cells with a gene-editing payload using a gas-propelled atomized spray from an isotonic aqueous alcohol-containing delivery solution, implemented in a system with differential pressure. The method and system further include delivery of stop solution and culture medium as part of the overall workflow.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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