Methods and products for transfecting cells

• US 9,605,278 B2
• Filed: 07/28/2016
• Issued: 03/28/2017
• Est. Priority Date: 12/05/2011
• Status: Active Grant

• Current Assignee: Factor Bioscience Inc
• Original Assignee: Factor Bioscience Inc
• Inventors: Angel, Matthew, Rohde, Christopher
• Primary Examiner(s): Noble, Marcia S
• Application Number:
  US15/222,453
• Publication Number:
  US 20160333378A1
• Time in Patent Office: 243 Days
• US Class Current: 1/1
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/18 for HIV
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/06 Antianaemics
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C08G 77/08 characterised by the catalysts used
• CPC Class Codes: C08K 5/5399 Phosphorus bound to nitrogen
• CPC Class Codes: C08L 83/04 : Polysiloxanes
• CPC Class Codes: C12N 15/87 : Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
• CPC Class Codes: C12N 15/907 : in mammalian cells
• CPC Class Codes: C12N 2500/25 : Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/40 : Nucleotides, nucleosides or bases
• CPC Class Codes: C12N 2500/44 : Thiols, e.g. mercaptoethanol
• CPC Class Codes: C12N 2501/115 : Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/155 : Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2501/165 : Vascular endothelial growth factor [VEGF]
• CPC Class Codes: C12N 2501/2303 : Interleukin-3 (IL-3)
• CPC Class Codes: C12N 2501/26 : Flt-3 ligand (CD135L, flk-2 ligand)
• CPC Class Codes: C12N 2501/91 : Heparin
• CPC Class Codes: C12N 2501/998 : Proteins not provided for elsewhere
• CPC Class Codes: C12N 2506/09 : from epidermal cells, from skin cells, from oral mucosa cells
• CPC Class Codes: C12N 2510/00 : Genetically modified cells
• CPC Class Codes: C12N 2800/80 : Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites
• CPC Class Codes: C12N 5/0647 : Haematopoietic stem cells; Uncommitted or multipotent progenitors
• CPC Class Codes: C12N 5/0657 : Cardiomyocytes; Heart cells
• CPC Class Codes: C12N 5/0696 : Artificially induced pluripotent stem cells, e.g. iPS
• CPC Class Codes: C12N 9/16 : acting on ester bonds (3.1)
• CPC Class Codes: C12N 9/22 : Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113
• CPC Class Codes: C12P 21/00 : Preparation of peptides or proteins single cell protein C12N1/00
• CPC Class Codes: C12Y 301/21 : Endodeoxyribonucleases producing 5'-phosphomonoesters (3.1.21)
• CPC Class Codes: H01L 31/048 : Encapsulation of modules
• CPC Class Codes: Y02E 10/50 : Photovoltaic [PV] energy
• CPC Class Codes: Y02E 10/52 : PV systems with concentrators

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.

Claims

1. 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 double-strand break in the DNA of the hematopoietic cell; and

a double-strand break in its DNA, the double-strand break being caused by the gene-editing protein and reducing the function of one or more of CCR5 and CXCR4.

2. The therapeutic composition of claim 1, wherein the gene-editing protein is selected from a TALEN and a zinc finger nuclease.

3. The therapeutic composition of claim 1, 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.

4. The therapeutic composition of claim 1, wherein the double-strand break is within about 5,000,000 bases of the transcription start site of the CCR5 or CXCR4 gene.

5. The therapeutic composition of claim 1, wherein the hematopoietic cell is a hematopoietic stem cell.

6. The therapeutic composition of claim 1, wherein the hematopoietic cell is a white blood cell.