Methods and compositions to increase somatic cell nuclear transfer (SCNT) efficiency by removing histone H3-lysine trimethylation

Patent metadata

Field	Value
Publication number	US-11390885-B2
Application number	US-201515511348-A
Country	US
Kind code	B2
Filing date	Sep 15, 2015
Priority date	Sep 15, 2014
Publication date	Jul 19, 2022
Grant date	Jul 19, 2022

Abstract

The present invention provides methods and compostions to improve the efficiency of somatic cell nuclear transfer (SCNT) and the consequent production of nuclear transfer ESC (ntESC) and transgenic cells and/or non-human animals. More specifically, the present invention relates to the discovery that trimethylation of Histone H3-Lysine 9 (H3K9me3) in reprogramming resistant regions (RRRs) in the nuclear genetic material of donor somatic cells prevents efficient somatic cell nuclear reprogramming or SCNT. The present invention provide methods and compositions to decrease H3K9me3 in methods to improve efficacy of SCNT by exogenous or overexpression of the demethylase Kdm4 family and/or inhibiting methylation of H3K9me3 by inhibiting the histone methyltransferases Suv39h1 and/or Suv39h2.

First claim

The invention claimed is:

1. A composition comprising: (i) a non-human mammalian Somatic Cell Nuclear Transfer (SCNT) embryo comprising a nucleus obtained from a terminally differentiated donor non-human mammalian somatic cell, (ii) a recipient non-human mammalian oocyte comprising a nucleus obtained from a terminally differentiated donor non-human mammalian somatic cell, or (iii) a non-human mammalian blastocyst developed from a non-human mammalian SCNT embryo comprising a nucleus obtained from a terminally differentiated donor non-human mammalian somatic cell, each comprising an exogenous mRNA polynucleotide encoding a Kdm4 histone demethylase and overexpressing the Kdm4 histone demethylase, wherein said composition increases efficiency of pre-implantation and post-implantation development of SCNT embryos comprising the nucleus of (i), (ii), or (iii) relative to a control SCNT embryo.

2. The composition of claim 1 , wherein the exogenous mRNA polynucleotide encoding the Kdm4 histone demethylase is transcribed from a polynucleotide comprising a nucleic acid sequence selected from SEQ ID NOs: 1-8.

3. The composition of claim 1 , wherein the non-human mammalian SCNT embryo is at the 1-cell or 2-cell stage.

4. The composition of claim 1 , wherein the recipient non-human mammalian oocyte is an enucleated recipient non-human mammalian oocyte.

5. The composition of claim 1 , wherein the recipient non-human mammalian oocyte is a recipient non-human mammalian oocyte.

6. The composition of claim 1 , wherein the non-human mammal is a domestic or commercial animal, a companion animal or pet, or from a mammalian species which is near extinction.

7. The composition of claim 1 , wherein the recipient non-human mammalian oocyte is from a non-human mammal.

8. The composition of claim 7 , wherein the non-human mammal is a domestic or commercial animal, a companion animal or pet, or from a mammalian species which is near extinction.

9. The composition of claim 1 , wherein the donor nucleus and/or the SCNT embryo comprises an inhibitor of one or more of Suv39h1, Suv39h2, or Setdb1 methyl transferase.

Assignees

• Childrens Medical Center

Inventors

• Zhang Yi
• Matoba Shogo

Classifications

• A01K2227/105

  Murine · CPC title

• C12Y201/01043

  Histone-lysine N-methyltransferase (2.1.1.43) · CPC title

• C12N15/877 Primary

  Techniques for producing new mammalian cloned embryos · CPC title

• A01K67/0273 Primary

  Cloned vertebrates · CPC title

• C12N15/1137

  against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138) · CPC title