Generation of healthy mice from gene-corrected disease-specific induced pluripotent stem cells.
Cast your vote
You can rate an item by clicking the amount of stars they wish to award to this item.
When enough users have cast their vote on this item, the average rating will also be shown.
Your vote was cast
Thank you for your feedback
Thank you for your feedback
Sharma, Amar Deep
Han, Dong Wook
Schöler, Hans R
MetadataShow full item record
AbstractUsing the murine model of tyrosinemia type 1 (fumarylacetoacetate hydrolase [FAH] deficiency; FAH⁻/⁻ mice) as a paradigm for orphan disorders, such as hereditary metabolic liver diseases, we evaluated fibroblast-derived FAH⁻/⁻-induced pluripotent stem cells (iPS cells) as targets for gene correction in combination with the tetraploid embryo complementation method. First, after characterizing the FAH⁻/⁻ iPS cell lines, we aggregated FAH⁻/⁻-iPS cells with tetraploid embryos and obtained entirely FAH⁻/⁻-iPS cell-derived mice that were viable and exhibited the phenotype of the founding FAH⁻/⁻ mice. Then, we transduced FAH cDNA into the FAH⁻/⁻-iPS cells using a third-generation lentiviral vector to generate gene-corrected iPS cells. We could not detect any chromosomal alterations in these cells by high-resolution array CGH analysis, and after their aggregation with tetraploid embryos, we obtained fully iPS cell-derived healthy mice with an astonishing high efficiency for full-term development of up to 63.3%. The gene correction was validated functionally by the long-term survival and expansion of FAH-positive cells of these mice after withdrawal of the rescuing drug NTBC (2-(2-nitro-4-fluoromethylbenzoyl)-1,3-cyclohexanedione). Furthermore, our results demonstrate that both a liver-specific promoter (transthyretin, TTR)-driven FAH transgene and a strong viral promoter (from spleen focus-forming virus, SFFV)-driven FAH transgene rescued the FAH-deficiency phenotypes in the mice derived from the respective gene-corrected iPS cells. In conclusion, our data demonstrate that a lentiviral gene repair strategy does not abrogate the full pluripotent potential of fibroblast-derived iPS cells, and genetic manipulation of iPS cells in combination with tetraploid embryo aggregation provides a practical and rapid approach to evaluate the efficacy of gene correction of human diseases in mouse models.
CitationGeneration of healthy mice from gene-corrected disease-specific induced pluripotent stem cells. 2011, 9 (7):e1001099 PLoS Biol.
AffiliationMax-Planck-Institute for Molecular Biomedicine, Münster, Germany.
The following license files are associated with this item:
- Curative ex vivo liver-directed gene therapy in a pig model of hereditary tyrosinemia type 1.
- Authors: Hickey RD, Mao SA, Glorioso J, Elgilani F, Amiot B, Chen H, Rinaldo P, Marler R, Jiang H, DeGrado TR, Suksanpaisan L, O'Connor MK, Freeman BL, Ibrahim SH, Peng KW, Harding CO, Ho CS, Grompe M, Ikeda Y, Lillegard JB, Russell SJ, Nyberg SL
- Issue date: 2016 Jul 27
- Grown up mice from gene-corrected iPS cells.
- Authors: Laidman J
- Issue date: 2011 Nov 11
- Renal proximal tubular cells acquire resistance to cell death stimuli in mice with hereditary tyrosinemia type 1.
- Authors: Luijerink MC, van Beurden EA, Malingré HE, Jacobs SM, Grompe M, Klomp LW, Berger R, van den Berg IE
- Issue date: 2004 Sep
- High volume naked DNA tail-vein injection restores liver function in Fah-knock out mice.
- Authors: Eggenhofer E, Doenecke A, Renner P, Slowik P, Piso P, Geissler EK, Schlitt HJ, Dahlke MH, Popp FC
- Issue date: 2010 May
- Kidneys of mice with hereditary tyrosinemia type I are extremely sensitive to cytotoxicity.
- Authors: Jacobs SM, van Beurden DH, Klomp LW, Berger R, van den Berg IE
- Issue date: 2006 Mar