GENERATING GABAergic NEURONS IN BRAINS

Abstract

This document provides methods and materials for generating GABAergic neurons in brains. For example, methods and materials for using nucleic acid encoding a NeuroD1 polypeptide and nucleic acid encoding a Dlx2 polypeptide to trigger glial cells (e.g., NG2 glial cells or astrocytes) within the brain (e.g., striatum) into forming GABAergic neurons (e.g., neurons resembling medium spiny neurons such as DARPP32-positive GABAergic neurons) that are functionally integrated into the brain of a living mammal (e.g., a human) are provided.

1 . (canceled) 
     
     
         2 . A composition for converting glial cells to GABAergic neurons in a striatum of a living mammal's brain, wherein said composition comprises a nucleic acid vector comprising a nucleic acid sequence encoding a neurogenic differentiation 1 (NeuroD1) polypeptide and a nucleic acid sequence encoding a distal-less homeobox 2 (Dlx2) polypeptide. 
     
     
         3 . The composition of  claim 2 , wherein said nucleic acid vector is a viral vector. 
     
     
         4 . The composition of  claim 3 , wherein said viral vector is an adeno-associated viral vector. 
     
     
         5 . The composition of  claim 2 , wherein said nucleic acid sequence encoding said NeuroD1 polypeptide or said nucleic acid sequence encoding said Dlx2 polypeptide is operably linked to a promoter sequence, wherein said promoter sequence is a constitutive promoter sequence. 
     
     
         6 . The composition of  claim 5 , wherein said constitutive promoter sequence is selected from the group consisting of a NG2 promoter sequence, a GFAP promoter sequence, an EF1a promoter sequence, a CMV promoter sequence, an Aldh1L1 promoter sequence, and a CAG promoter sequence. 
     
     
         7 . The composition of  claim 2 , wherein said nucleic acid sequence encoding said NeuroD1 polypeptide or said nucleic acid sequence encoding said Dlx2 polypeptide is operably linked to a promoter sequence, wherein said promoter sequence is a glial-specific promoter sequence. 
     
     
         8 . The composition of  claim 7 , wherein said glial-specific promoter sequence is selected from the group consisting of a NG2 promoter sequence, a GFAP promoter sequence, an A1dh1L1 promoter sequence, and an Olig2 promoter sequence. 
     
     
         9 . The composition of  claim 2 , wherein said mammal is diagnosed with Huntington's disease. 
     
     
         10 . The composition of  claim 9 , wherein said mammal is a human. 
     
     
         11 . The composition of  claim 2 , wherein said glial cells are NG2 glial cells. 
     
     
         12 . The composition of  claim 2 , wherein said GABAergic neurons are DARPP32-positive. 
     
     
         13 . The composition of  claim 2 , wherein said NeuroD1 polypeptide is a human NeuroD1 polypeptide comprising an amino acid sequence of SEQ ID NO: 1. 
     
     
         14 . The composition of  claim 2 , wherein said Dlx2 polypeptide is a human Dlx2 peptide comprising an amino acid sequence of SEQ ID NO: 2.