Brain-specific enhancers for cell-based therapy

Patent metadata

Field	Value
Publication number	US-9952216-B2
Application number	US-201414512306-A
Country	US
Kind code	B2
Filing date	Oct 10, 2014
Priority date	Apr 10, 2012
Publication date	Apr 24, 2018
Grant date	Apr 24, 2018

Abstract

Herein are described a set of novel specific human enhancers for specific forebrain cell types used to study and select for human neural progenitor cells. This approach enables the ability to generate interneurons from human ES, iPS and iN cells, making them available for human transplantation and for molecular/cellular analyzes. These approaches are also directly applicable to generating other neuronal cell types, such as cortical and striatal projection neurons, which have implications for many human diseases.

First claim

What is claimed is:

1. An isolated polynucleotide comprising an enhancer comprising the sequence of SEQ ID NO: 120, and further comprising a heterologous inducible promoter and a reporter gene, wherein the enhancer and the heterologous inducible promoter are operably linked to the reporter gene.

2. The isolated polynucleotide of claim 1 further comprising a gene that is capable of inducing pluripotency.

3. A vector comprising an isolated polynucleotide of claim 1 .

4. An expression cassette incorporating the isolated polynucleotide of claim 1 .

5. A non-human cell comprising the expression cassette of claim 4 .

6. A vector comprising an isolated polynucleotide of claim 2 .

7. A set of vectors for labeling subregions of a mouse forebrain, comprising the vector of claim 3 , and a second vector comprising an enhancer selected from the group consisting of SEQ ID NOS: 83, 84, 99-104, 106-108, 110-118, 121-128, and 144-145.

8. A method for detecting oligodendrocyte progenitors comprising: (1) providing the vector of claim 3 ; (2) transfecting an embryonic stem cell with the vector; (3) directing neuronal differentiation of the transfected embryonic stem cell; and (4) detecting oligodendrocyte progenitors by detecting reporter gene expression.

9. A method for detecting and isolating oligodendrocyte progenitors comprising (1) providing the vector of claim 3 ; (2) transfecting an embryonic stem cell with the vector; (3) directing neuronal differentiation of the transfected embryonic stem cell; (4) detecting oligodendrocyte progenitors by detecting reporter gene expression; and (5) isolating the oligodendrocyte progenitors expressing the reporter gene.

10. A method for driving reporter gene expression in medial ganglionic eminence (MGE) ventricular zone (VZ) or subventricular zone (SVZ), or preoptic area (POA) ventricular zone (VZ) or subventricular zone (SVZ) of a mouse, comprising (1) providing the vector of claim 3 ; (2) using the vector to generate a transgenic mouse; and (3) detecting expression of the reporter gene in the MGE or POA of the transgenic mouse.

Assignees

• Visel Axel
• Rubenstein John L R
• Chen Ying Jiun Jasmine
• Pennacchio Len A
• Vogt Daniel
• Nicholas Cory
• Kriegstein Arnold
• Univ California

Inventors

• Visel Axel
• Rubenstein John L R
• Chen Ying-Jiun (Jasmine)
• Pennacchio Len A
• Vogt Daniel
• Nicholas Cory
• Kriegstein Arnold

Classifications

• C12N15/86

  Viral vectors · CPC title

• C12N15/85 Primary

  for animal cells · CPC title

• G01N33/56966 Primary

  Animal cells · CPC title

• A61K35/545

  Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells · CPC title

• G01N33/5014

  for testing toxicity · CPC title