Methods and products for transfecting cells

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.

1 .- 61 . (canceled) 
     
     
         62 . A therapeutic composition, comprising a gene-edited hematopoietic cell, wherein the gene-edited hematopoietic cell comprises:
 an in vitro transcribed synthetic RNA molecule encoding a gene-editing protein, the gene-editing protein comprising a DNA-binding domain and a nuclease catalytic domain that causes a single-strand or double-strand break in the DNA of the hematopoietic cell; and   a single-strand or double-strand break in its DNA, the single-strand or double-strand break being caused by the gene-editing protein and reducing the function of a HBB gene.   
     
     
         63 . The therapeutic composition of  claim 62 , wherein the gene-editing protein is selected from a TALEN and a zinc finger nuclease. 
     
     
         64 . The therapeutic composition of  claim 62 , wherein the in vitro transcribed synthetic RNA molecule further comprises one or more of a 5′-cap, a 5′-cap 1 structure, and a 3′-poly(A) tail. 
     
     
         65 . The therapeutic composition of  claim 62 , wherein the double-strand break is within about 5,000,000 bases of the transcription start site of the HBB gene. 
     
     
         66 . The therapeutic composition of  claim 62 , wherein the hematopoietic cell is a hematopoietic stem cell. 
     
     
         67 . The therapeutic composition of  claim 62 , wherein the hematopoietic cell is a white blood cell.