Methods for phototransfecting nucleic acids into live cells
AssigneeThe Trustees of the University of Pennsylvania
InventorsJames Eberwine, Philip G. Haydon, Jai-Yoon Sul, Hajime Takano, Chia-Wen Kitty Wu, Fanyi Zeng
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Abstract
The present invention includes methods for transferring a multigenic phenotype to a cell by transfecting, preferably by phototransfection, and locally transfecting a cell or a cellular process with a laser while the cell is bathed in a fluid medium comprising two or more nucleic acids, thereby introducing the nucleic acid into the interior of the cell. Expression of the nucleic acids results in a multigenic phenotype in the transfected cell.
Claims15
A method of transferring a phenotype of a first cell to a second cell, said method comprising
- transfecting the second cell with an mRNA transcriptome of the first cell a first time, wherein transfecting the second cell comprises irradiating the second cell with a laser, wherein said second cell is bathed in a fluid medium comprising the mRNA transcriptome of the first cell, wherein said second cell is irradiated ex vivo or in vitro, wherein said laser porates a cellular membrane of said second cell and said mRNA transcriptome enters said second cell, wherein said mRNA transcriptome is functional in the second cell, and
- transfecting the second cell with an mRNA transcriptome of the first cell at least a second time, wherein transfecting the second cell comprises irradiating the second cell with a laser, wherein said second cell is bathed in a fluid medium comprising the mRNA transcriptome of the first cell, wherein said second cell is irradiated ex vivo or in vitro, wherein said laser porates a cellular membrane of said second cell and said mRNA transcriptome enters said second cell wherein said mRNA transcriptome is functional in the second cell,
- thereby initiating a change in physiology and morphology of the second cell, wherein the change in physiology and morphology yields a phenotype of the second cell that is indicative of the first cell
- wherein the first cell and second cell are selected from the group consisting of:
- a) wherein the second cell is a fibroblast, and wherein the first cell is selected from the group consisting of an astrocyte, a cardiomyocyte, and a stem cell;
- b) wherein the second cell is a neuron, and wherein the first cell is selected from the group consisting of an astrocyte and a cardiomyocyte;
- and
- c) wherein the second cell is an astrocyte and wherein the first cell is a cardiomyocyte.
Dependencies are inferred from the claim text.