Methods for enhancing genome stability and telomere elongation in embryonic stem cells

• US 9,012,223 B2
• Filed: 04/23/2014
• Issued: 04/21/2015
• Est. Priority Date: 09/04/2009
• Status: Active Grant

• Current Assignee: United States Department Of Health And Human Services
• Original Assignee: United States Department Of Health And Human Services
• Inventors: Zalzman, Michal, Sharova, Lioudmila V., Ko, Minoru S. H.
• Primary Examiner(s): BERTOGLIO, VALARIE E
• Application Number:
  US14/259,600
• Publication Number:
  US 20140234969A1
• Time in Patent Office: 363 Days
• Field of Search: 435/455
• US Class Current: 435/455
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 39/00 General protective or antinoxious agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/4705 stimulating, promoting or activating activity
• CPC Class Codes: C12N 15/00 Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor mutants or genetically engineered microorganisms, per se C12N1/00, C12N5/00, C12N7/00; new plants per se A01H; plant reproduction by tissue culture techniques A01H4/00; new animals per se A01K67/00; use of medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases, gene therapy A61K48/00
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2501/00 Active agents used in cell culture processes, e.g. differentation
• CPC Class Codes: C12N 2501/40 Regulators of development
• CPC Class Codes: C12N 2501/60 Transcription factors
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/00 Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor; plant reproduction by tissue culture techniques A01H4/00
• CPC Class Codes: C12N 5/0606 Pluripotent embryonic cells, e.g. embryonic stem cells [ES] embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696

Abstract

The disclosure provides methods for increasing genome stability of an embryonic stem (ES) cell or induced pluripotent stem (iPS) cell, increasing telomere length in an ES or iPS cell, or both, for example by contacting an ES or iPS cell with an agent that increases expression of Zscan4 in the cell. Methods for increasing genome stability or increasing telomere length in a population of ES or iPS cells are provided, for example by selecting Zscan4⁺ ES or iPS cells from the population of ES or iPS cells (which can include both Zscan4⁺ and Zscan4⁻ ES or iPS cells). Therapeutic methods of using ES or iPS cells expressing Zscan4 are also provided. Further provided are methods of treating cancer by administering a Zscan4 polynucleotide or Zscan4 polypeptide. Also provided are methods of inducing differentiation of isolated ES or iPS cells into germ cells.

Claims

1. A method for increasing genome stability of an isolated mouse induced pluripotent stem (iPS) cell;

or increasing telomere length in an isolated mouse iPS cell;

or both, comprising transfecting an isolated nucleic acid molecule encoding mouse Zscan4c into the iPS cell under conditions sufficient to allow for expression of mouse Zscan4c in the iPS cell.

2. The method of claim 1, wherein the isolated nucleic acid molecule comprises a vector.

3. The method of claim 2, wherein the vector is a viral vector.

4. The method of claim 2, wherein the vector is a plasmid vector.

5. The method of claim 2, wherein the vector encodes mouse Zscan4c operably linked to a promoter.

6. The method of claim 5, wherein the promoter is a constitutive promoter.

7. The method of claim 5, wherein the promoter is an inducible promoter.

8. The method of claim 1, wherein the nucleotide sequence of mouse Zscan4c is at least 95% identical to the nucleotide sequence of SEQ ID NO:

28.

9. The method of claim 1, wherein the nucleotide sequence of mouse Zscan4c comprises the nucleotide sequence of SEQ ID NO:

28.

10. The method of claim 1, wherein the nucleotide sequence of mouse Zscan4c consists of the nucleotide sequence of SEQ ID NO:

28.

11. The method of claim 1, wherein expression of mouse Zscan4c is transient.

12. The method of claim 1, wherein the isolated nucleic acid molecule encoding mouse Zscan4c is integrated into the genome of the iPS cell.