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Successful Derivation of an Induced Pluripotent Stem Cell Line from a Genetically Nonpermissive Enhanced Green Fluorescent Protein-Transgenic FVB/N Mouse Strain

Abbey, Deepti and Singh, Gurbind and Verma, Isha and Derebail, Suchitra and Kolkundkar, Udaykumar and Chandrashekar, Darshan S and Acharya, Kshitish K and Vemuri, Mohan C and Seshagiri, Polani B (2019) Successful Derivation of an Induced Pluripotent Stem Cell Line from a Genetically Nonpermissive Enhanced Green Fluorescent Protein-Transgenic FVB/N Mouse Strain. In: CELLULAR REPROGRAMMING, 21 (5). pp. 270-284.

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Official URL: https://dx.doi.org/10.1089/cell.2019.0019

Abstract

The embryonic stem cell line derivation from nonpermissive mouse strains is a challenging and highly inefficient process. The cellular reprogramming strategy provides an alternative route for generating pluripotent stem cell (PSC) lines from such strains. In this study, we successfully derived an enhanced green fluorescent protein (EGFP)-transgenic ``N9'' induced pluripotent stem cell (iPS cell, iPSC) line from the FVB/N strain-derived mouse embryonic fibroblasts (MEFs). The exposure of MEFs to human OCT4, SOX2, KLF4, and c-MYC (OSKM) transgenes via lentiviral transduction resulted in complete reprogramming. The N9 iPS cell line demonstrated all the criteria of a typical mouse PSC line, including normal colony morphology and karyotype (40,XY), high replication and propagation efficiencies, expression of the pluripotency-associated genes, spontaneous differentiation to three germ lineage-derived cell types, and robust potential of chimeric blastocyst formation. Taken together, using human OSKM genes for transduction, we report, for the first time, the successful derivation of an EGFP-expressing iPS cell line from a genetically nonpermissive transgenic FVB/N mouse. This cell line could provide opportunities for designing protocols for efficient derivation of PSC lines from other nonpermissive strains and developing mouse models of human diseases.

Item Type: Journal Article
Additional Information: Copyright of this article belongs to MARY ANN LIEBERT, INC
Keywords: induced pluripotent stem cells; transgenic; nonpermissive; enhanced green fluorescent protein; differentiation; reprogramming
Department/Centre: Division of Biological Sciences > Molecular Reproduction, Development & Genetics
Depositing User: Id for Latest eprints
Date Deposited: 29 Jan 2020 11:11
Last Modified: 29 Jan 2020 11:11
URI: http://eprints.iisc.ac.in/id/eprint/63850

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