Isolated mouse or human cell having an exogenous transgene in an endogenous albumin gene

• US 9,255,250 B2
• Filed: 12/04/2013
• Issued: 02/09/2016
• Est. Priority Date: 12/05/2012
• Status: Active Grant

• Current Assignee: Children's Hospital of Philadelphia (Children's Hospital of Philadelphia Foundation), Sangamo Biosciences, Inc.
• Original Assignee: Children's Hospital of Philadelphia (Children's Hospital of Philadelphia Foundation), Sangamo Biosciences, Inc.
• Inventors: Gregory, Philip D., High, Katherine A.
• Primary Examiner(s): WILSON, MICHAEL C
• Application Number:
  US14/096,788
• Publication Number:
  US 20140155468A1
• Time in Patent Office: 797 Days
• Field of Search: 435/435, 435/455, 435/325
• 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: A61K 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 13/02 of urine or of the urinary tract, e.g. urine acidifiers
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: C12N 5/10 Cells modified by introduction of foreign genetic material

Abstract

Cells and methods of using these cells for expressing a transgene expressing a protein that is aberrantly expressed in a metabolic disorders from a safe harbor locus.

Claims

1. An isolated mouse or human cell comprising an exogenous transgene encoding a protein, wherein the exogenous transgene is integrated site-specifically using a nuclease into an endogenous albumin gene, wherein the protein is a functional version of a protein selected from the group consisting of a methylmalonic aciduria CbIA Type (MMAA) protein, a methylmalonic aciduria CbIB Type (MMAB) protein, a methylmalonic aciduria CbIC Type (MMADHC) protein, a 5-Methyltetrahydrofolate-Homocysteine Methyltransferase Reductase (MTRR) protein, a lysosomal membrane protein domain (LMBRD 1) protein, a 5-Methyltetrahydrofolate-Homocysteine Methyltransferase (MTR) protein, a propionyl-CoA protein, a glucose-6-phosphate transporter (G6PT) protein, a glucose-6-phosphatase (G6Pase) protein, a low density lipoprotein receptor (LDLR) protein, a low density lipoprotein receptor adaptor protein 1 (LDLRAP-1 protein), N-acetylglutamate synthetase (NAGS) protein, a carbamoyl phosphate synthetase 1 (CPS1) protein, an ornthine transcarbamylase (OTC) protein, an argininosuccinic acid synthetase (ASS) protein, an argininosuccinase acid lyase (ASL) protein, an arginase (ARG1) protein, a solute carrier family 25 protein, a UDP glucuronosyltransferase 1 family, polypeptide A1 (UGT1A1) protein, a fumarylacetoacetate hydrolyase (FAH), an alanine-glyoxylate aminotransferase (AGXT) protein, a glyoxylate reductase/hydroxypyruvate reductase (GRHPR) protein, an APTase Cu(2+) transporting beta (ATP7B) protein, a phenylalanine hydroxylase (PAH) protein and a lipoprotein lyase (LPL) protein.

2. The cell of claim 1, wherein the cell is selected from the group consisting of a hepatic cell, an induced pluripotent stem cell (iPSC), a hematopoietic stem cell, a hepatic cell, a hepatic stem cell, and a red blood precursor cell.

3. The cell of claim 1, wherein expression of the transgene is driven by an endogenous albumin promoter.

4. The cell of claim 1, wherein the transgene is operably linked to a promoter sequence that drives expression of the transgene.

5. A method of generating a genetically modified cell according to claim 1, the method comprising administering a nuclease and a vector encoding the transgene to the cell such that the transgene is integrated into the endogenous albumin gene.