Methods and products for transfecting cells
AssigneeFactor Bioscience Inc.
InventorsMatthew Angel, Christopher Rohde
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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.
Claims32
A method for altering a target DNA sequence in a cell, comprising:
- (a) providing a human somatic cell comprising a target DNA sequence;
- (b) culturing the cell;
- and
- (c) transfecting the cell with a plurality of synthetic RNA molecules, wherein the synthetic RNA molecules include:
- i. a first synthetic RNA molecule encoding a first fusion protein comprising a DNA-binding domain and a catalytic domain of a nuclease;
- and
- ii. a second synthetic RNA molecule encoding a second fusion protein comprising a DNA-binding domain and a catalytic domain of a nuclease;
- wherein:
- the first fusion protein and the second fusion protein are independently a transcription activator-like effector nuclease (TALEN);
- and
- the transfecting results in the cell expressing the first fusion protein and the second fusion protein to result in a single-strand break or a double-strand break.
A method for altering a target DNA sequence in a cell, comprising:
- (a) providing a human somatic cell comprising a target DNA sequence,
- (b) culturing the cell;
- (c) transfecting the cell with a plurality of synthetic RNA molecules, wherein the synthetic RNA molecules include:
- i. a first synthetic RNA molecule encoding a first fusion protein comprising a DNA-binding domain and a catalytic domain of a nuclease;
- and
- ii. a second synthetic RNA molecule encoding a second fusion protein comprising a DNA-binding domain and a catalytic domain of a nuclease;
- wherein the first fusion protein and the second fusion protein are independently a transcription activator-like effector nuclease (TALEN);
- and wherein the transfecting results in the cell expressing the first fusion protein and the second fusion protein to result in a single-strand break or a double-strand break in the target DNA sequence;
- and
- (d) transfecting the cell with a DNA repair template comprising a sequence for insertion and one or more regions of homology to the DNA of the cell, wherein the one or more regions of homology comprise regions upstream and/or downstream of the single-strand break or the double-strand break, to result in insertion of the sequence in the region of the single-strand break or the double-strand break.
Dependencies are inferred from the claim text.