METHODS FOR MODULATING EMBRYONIC STEM CELL DIFFERENTIATION

• US 20140120579A1
• Filed: 12/30/2013
• Published: 05/01/2014
• Est. Priority Date: 03/26/2007
• Status: Abandoned Application

• Current Assignee: United States Department Of Health And Human Services (Government of the United States of America)
• Original Assignee: The United States of America As Represented By The Secretary of Agriculture
• Inventors: KO, MINORU S.H., FALCO, GEPPINO, LEE, SUNG-LIM, MONTI, MANUELA, STANGHELLINI, ILARIA
• Application Number:
  US14/143,568
• Publication Number:
  US 20140120579A1
• Time in Patent Office: Days
• US Class Current: 435/69.1
• CPC Class Codes: A01K 2217/052 inducing gain of function
• CPC Class Codes: A01K 2217/15 Animals comprising multiple alterations of the genome, by transgenesis or homologous recombination, e.g. obtained by cross-breeding
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/0393 Animal model comprising a reporter system for screening tests
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: C07K 14/47 from mammals
• CPC Class Codes: C12N 15/113 Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing when used in plants C12N15/8218
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 2310/111 spanning the whole gene, or a large part of it
• CPC Class Codes: C12N 2310/14 interfering N.A.
• CPC Class Codes: C12N 2310/53 partially self-complementary or closed
• CPC Class Codes: C12N 2501/40 Regulators of development
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2830/008 cell type or tissue specific enhancer/promoter combination
• CPC Class Codes: C12N 2830/85 mammalian
• 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
• CPC Class Codes: C12N 5/0611 Primordial germ cells, e.g. embryonic germ cells [EG]

Abstract

Described herein is Zscan4, a gene exhibiting 2-cell embryonic stage and embryonic stem cell specific expression. Identification of nine Zscan4 co-expressed genes is also described Inhibition of Zscan4 expression inhibits the 2-cell to 4-cell embryonic transition and prevents blastocyst implantation, expansion and outgrowth. Provided herein are methods of inhibiting differentiation of a stem cell, promoting blastocyst outgrowth of embryonic stem cells and identifying a subpopulation of stem cells expressing Zscan4. Further described is the identification of Trim43 as a gene exhibiting morula-specific expression. Also provided are isolated expression vectors comprising a Zscan4 promoter, or a Trim43 promoter operably linked to a heterologous polypeptide and uses thereof. Further provided are transgenic animals comprising transgenes encoding marker proteins operably linked to Zscan4 and Trim43 promoters.

Claims

1. An in vitro method of increasing the expression of a Zscan4 protein in an embryonic stem cell comprising introducing a nucleic acid encoding the Zscan4 protein into the embryonic stem cell.

2. The method of claim 1, wherein the nucleic acid is operably linked to a promoter.

3. The method of claim 2, wherein the nucleic acid encoding the Zscan4 protein comprises SEQ ID NO:

60.

4. The method of claim 2, wherein the nucleic acid encoding the Zscan4 protein is at least 95% identical to SEQ ID NO:

19, SEQ ID NO;

21, SEQ ID NO;

25, or SEQ ID NO;

29.

5. The method of claim 1, further comprising a vector comprising the nucleic acid encoding the Zscan4 protein.