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Methods of using ZSCAN4 for rejuvenating human cells

A cell, object technology applied in the field of rejuvenation of human cells using ZSCAN4

Inactive Publication Date: 2016-01-13
ELIXIRGEN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although it has been shown that the human genome also contains the ZSCAN4 gene, Falco et al., DevBiol 307:539-550, 2007; Zalzman et al., Nature 464:858-863, 2010; PCT Publication Nos. WO2008 / 118957, WO2011 / 02880, WO2012 / 103235, None of WO2012 / 129342, WO2012 / 158561, and WO2012158564; or U.S. Patent Application Publication Nos. US2010 / 0105043, US2012 / 0129161, and US2012 / 0156305 provided experiments showing that Zscan4 expression in human cells resulted in the same effects as in murine embryonic cells support

Method used

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  • Methods of using ZSCAN4 for rejuvenating human cells
  • Methods of using ZSCAN4 for rejuvenating human cells
  • Methods of using ZSCAN4 for rejuvenating human cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0368] Example 1: Zscan4 Expression Corrects Chromosomal Abnormalities in Mouse Embryonic Stem Cells

[0369] This example describes the discovery that expression of Zscan4, either by synthetic mRNA encoding Zscan4 or by a Sendai virus vector expressing Zscan4, can correct chromosomal abnormalities in mouse embryonic stem (ES). This example also demonstrates that synthetic mRNA encoding Zscan4 and Sendai virus vectors expressing Zscan4 can be used as biologics.

[0370] Materials and methods

[0371] cell culture

[0372]The MC1ZE murine ES cell line was previously reported (Amano et al., Nat Commun, 2013; 4:1966). MC1ZE cells were used as typical murine ES cells, which showed poor karyotype (ie, only about 20% of cells carried euploidy) due to long-term culture (passage number 33), and integration of exogenous genes. Cells were cultured at 37°C in 5% CO2 in complete ES medium as previously described (Amano et al., NatCommun, 2013; 4:1966.): DMEM (Gibco), 15% FBS (Atlanta...

Embodiment 2

[0389] Example 2: Effect of Zscan4 biological products on mouse embryonic stem cells

[0390] It has previously been shown that forced expression of exogenous murine Zscan4c from a plasmid vector integrated into the murine genome induces the expression of a unique set of genes including Tmem92, Stra8 and the endogenous Zscan4 gene (Amano et al., NatCommun, 2013; 4: 1966).

[0391] This example demonstrates that a Zscan4 biologic (eg, expressing Zscan4, either by a synthetic mRNA encoding Zscan4 or by a Sendai virus vector expressing Zscan4) exerts the same function in murine embryonic stem (ES) cells.

[0392] Materials and methods

[0393] cell culture

[0394]The MC1 murine embryonic stem (ES) cell line was previously used as a typical murine pluripotent stem cell to demonstrate the function of the exogenously introduced Zscan4c gene, which is integrated into the murine genome (Amano et al., NatCommun, 2013; 4:1966.) . MC1 cells were cultured at 37°C in 5% CO2 in compl...

Embodiment 3

[0405] Example 3: Effect of Zscan4 biological products on human induced pluripotent stem cells

[0406] The genes SERPINB4, DNMT3L, and DUX4 were identified as marker genes that were upregulated when the murine Zscan4c or human ZSCAN4 genes were overexpressed by a transgene-based gene expression system.

[0407] This example demonstrates that Zscan4 biologics (e.g., expressing Zscan4—either by synthetic mRNA encoding Zscan4 or by a Sendai virus vector expressing Zscan4) can be produced in a manner similar to that of the transgene-based Zscan4 overexpression system used in murine pluripotent stem cells. way to act on human iPS cells. This example also demonstrates that synthetic mRNA encoding human ZSCAN4 and Sendai virus vectors expressing human ZSCAN4 can be used as therapeutic biologics for the treatment of human pluripotent stem cells.

[0408] Materials and methods

[0409] cell culture

[0410] Human foreskin fibroblasts were cultured on mitotically inactivated murin...

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Abstract

The present disclosure relates to methods for increasing telomere length in one or more human cells and / or increasing genome stability of one or more human cells, for example by contacting one or more human cells with an agent that increases expression of Zscan4 in the one or more human cells. Methods of treating a subject in need of telomere lengthening, treating a disease or condition associated with a genomic and / or chromosome abnormality, of rejuvenating one or more human cells, of rejuvenating tissues or organs, and of rejuvenating a subject in need thereof, for example by contacting one or more human cells in the subject with an agent that increases expression of Zscan4, or by administering to a subject in need thereof, an agent that increases expression of Zscan4 are also provided.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of US Provisional Application No. 61 / 800,668, filed March 15, 2013, which is incorporated herein by reference in its entirety. [0003] Submit a sequence listing on an ASCII text file [0004] The following submission on ASCII text file is hereby incorporated by reference in its entirety: Sequence Listing in Computer Readable Form (CRF) (File Name: 699442000840SEQLIST.TXT, Date of Record: March 11, 2014, Size: 104KB) . technical field [0005] The present disclosure relates to methods of increasing telomere length and / or increasing genomic stability of one or more human cells in one or more human cells, for example, by combining one or more human cells with The one or more human cells are contacted with an agent that increases expression of Zscan4. The present disclosure further provides methods of treating a subject in need of telomere lengthening, methods of treating a disease or ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7088A61K48/00C12N15/11C12N15/09
CPCC12N2760/18841A61K48/005A61K31/7088A61K38/1709C07K14/47C12N15/113C12N2310/14C12N2310/531A61P1/00A61P1/02A61P1/04A61P1/16A61P1/18A61P11/00A61P13/00A61P13/02A61P13/08A61P13/10A61P13/12A61P15/00A61P17/00A61P17/02A61P17/06A61P17/14A61P19/00A61P19/02A61P21/00A61P21/02A61P21/04A61P25/00A61P25/14A61P25/16A61P25/28A61P25/32A61P27/02A61P27/16A61P29/00A61P35/00A61P35/02A61P37/02A61P37/06A61P37/08A61P43/00A61P5/00A61P5/38A61P7/00A61P7/02A61P7/06A61P9/00A61P9/10A61P3/10A61K38/57C12N15/63A61K48/00C12N15/86C12N2760/18843C12N15/09C12N2501/998C12N5/0606C12N5/0609C12N5/0696C12N2510/04C12Q1/6806C12Q1/6886C12Q2600/156C12N15/85A61K9/51A61K35/28A61K35/33A61K35/545C12N2320/31C12Q1/6883C12Q2600/136C12Q2600/158C12Q2521/107C12Q2521/327C12Q2537/159
Inventor M·S·H·科
Owner ELIXIRGEN
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