Methods and products for transfecting cells
First Claim
1. A method for producing a gene-edited, reprogrammed cell, comprising:
- (a) providing a differentiated cell;
(b) culturing the differentiated cell; and
(c) transfecting the differentiated cell with one or more synthetic RNA molecules,wherein the one or more synthetic RNA molecules include;
(i) at least one RNA molecule encoding one or more reprogramming factors, and(ii) at least one RNA molecule encoding one or more gene-editing proteins;
wherein the transfecting results in the cell expressing the one or more reprogramming factors and the one or more gene editing proteins to result in a gene-edited, reprogrammed cell;
wherein step (c) is performed without using irradiated human neonatal fibroblast feeder cells and occurs in the presence of a medium containing ingredients that support reprogramming of the cell to a less differentiated state.
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Abstract
The present invention relates in part to nucleic acids encoding proteins, nucleic acids containing non-canonical nucleotides, therapeutics comprising nucleic acids, methods, kits, and devices for inducing cells to express proteins, methods, kits, and devices for transfecting, gene editing, and reprogramming cells, and cells, organisms, and therapeutics produced using these methods, kits, and devices. Methods for inducing cells to express proteins and for reprogramming and gene-editing cells using RNA are disclosed. Methods for producing cells from patient samples, cells produced using these methods, and therapeutics comprising cells produced using these methods are also disclosed.
105 Citations
16 Claims
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1. A method for producing a gene-edited, reprogrammed cell, comprising:
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(a) providing a differentiated cell; (b) culturing the differentiated cell; and (c) transfecting the differentiated cell with one or more synthetic RNA molecules, wherein the one or more synthetic RNA molecules include; (i) at least one RNA molecule encoding one or more reprogramming factors, and (ii) at least one RNA molecule encoding one or more gene-editing proteins; wherein the transfecting results in the cell expressing the one or more reprogramming factors and the one or more gene editing proteins to result in a gene-edited, reprogrammed cell; wherein step (c) is performed without using irradiated human neonatal fibroblast feeder cells and occurs in the presence of a medium containing ingredients that support reprogramming of the cell to a less differentiated state. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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9. A method for producing a gene-edited, reprogrammed cell, comprising:
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(a) providing a non-pluripotent cell; (b) culturing the non-pluripotent cell; and (c) transfecting the non-pluripotent cell with one or more synthetic RNA molecules, wherein the one or more synthetic RNA molecules include; (i) at least one RNA molecule encoding one or more reprogramming factors, and (ii) at least one RNA molecule encoding one or more gene-editing proteins; wherein the transfecting results in the cell expressing the one or more reprogramming factors and the one or more gene editing proteins to result in a gene-edited, reprogrammed cell; wherein step (c) is performed without using irradiated human neonatal fibroblast feeder cells and occurs in the presence of a medium containing ingredients that support reprogramming of the cell. - View Dependent Claims (10, 11, 12, 13, 14, 15, 16)
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Specification