Proof-of-Concept Gene Editing for the Murine Model of Inducible Arginase-1 Deficiency

Abstract Arginase-1 deficiency in humans is a rare genetic disorder of metabolism resulting from a loss of arginase-1, leading to impaired ureagenesis, hyperargininemia and neurological deficits. Previously, we generated a tamoxifen-inducible arginase-1 deficient mouse model harboring a deletion of...

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Autores principales:	Yuan Yan Sin, Phillipe R. Price, Laurel L. Ballantyne, Colin D. Funk
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/eae6a3ed458b46789cceae268bf9e7a9
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spelling	oai:doaj.org-article:eae6a3ed458b46789cceae268bf9e7a92021-12-02T15:05:34ZProof-of-Concept Gene Editing for the Murine Model of Inducible Arginase-1 Deficiency10.1038/s41598-017-02927-22045-2322https://doaj.org/article/eae6a3ed458b46789cceae268bf9e7a92017-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-02927-2https://doaj.org/toc/2045-2322Abstract Arginase-1 deficiency in humans is a rare genetic disorder of metabolism resulting from a loss of arginase-1, leading to impaired ureagenesis, hyperargininemia and neurological deficits. Previously, we generated a tamoxifen-inducible arginase-1 deficient mouse model harboring a deletion of Arg1 exons 7 and 8 that leads to similar biochemical defects, along with a wasting phenotype and death within two weeks. Here, we report a strategy utilizing the Clustered, Regularly Interspaced, Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system in conjunction with piggyBac technology to target and reincorporate exons 7 and 8 at the specific Arg1 locus in attempts to restore the function of arginase-1 in induced pluripotent stem cell (iPSC)-derived hepatocyte-like cells (iHLCs) and macrophages in vitro. While successful gene targeted repair was achieved, minimal urea cycle function was observed in the targeted iHLCs compared to adult hepatocytes likely due to inadequate maturation of the cells. On the other hand, iPSC-derived macrophages expressed substantial amounts of “repaired” arginase. Our studies provide proof-of-concept for gene-editing at the Arg1 locus and highlight the challenges that lie ahead to restore sufficient liver-based urea cycle function in patients with urea cycle disorders.Yuan Yan SinPhillipe R. PriceLaurel L. BallantyneColin D. FunkNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-12 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yuan Yan Sin Phillipe R. Price Laurel L. Ballantyne Colin D. Funk Proof-of-Concept Gene Editing for the Murine Model of Inducible Arginase-1 Deficiency
description	Abstract Arginase-1 deficiency in humans is a rare genetic disorder of metabolism resulting from a loss of arginase-1, leading to impaired ureagenesis, hyperargininemia and neurological deficits. Previously, we generated a tamoxifen-inducible arginase-1 deficient mouse model harboring a deletion of Arg1 exons 7 and 8 that leads to similar biochemical defects, along with a wasting phenotype and death within two weeks. Here, we report a strategy utilizing the Clustered, Regularly Interspaced, Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system in conjunction with piggyBac technology to target and reincorporate exons 7 and 8 at the specific Arg1 locus in attempts to restore the function of arginase-1 in induced pluripotent stem cell (iPSC)-derived hepatocyte-like cells (iHLCs) and macrophages in vitro. While successful gene targeted repair was achieved, minimal urea cycle function was observed in the targeted iHLCs compared to adult hepatocytes likely due to inadequate maturation of the cells. On the other hand, iPSC-derived macrophages expressed substantial amounts of “repaired” arginase. Our studies provide proof-of-concept for gene-editing at the Arg1 locus and highlight the challenges that lie ahead to restore sufficient liver-based urea cycle function in patients with urea cycle disorders.
format	article
author	Yuan Yan Sin Phillipe R. Price Laurel L. Ballantyne Colin D. Funk
author_facet	Yuan Yan Sin Phillipe R. Price Laurel L. Ballantyne Colin D. Funk
author_sort	Yuan Yan Sin
title	Proof-of-Concept Gene Editing for the Murine Model of Inducible Arginase-1 Deficiency
title_short	Proof-of-Concept Gene Editing for the Murine Model of Inducible Arginase-1 Deficiency
title_full	Proof-of-Concept Gene Editing for the Murine Model of Inducible Arginase-1 Deficiency
title_fullStr	Proof-of-Concept Gene Editing for the Murine Model of Inducible Arginase-1 Deficiency
title_full_unstemmed	Proof-of-Concept Gene Editing for the Murine Model of Inducible Arginase-1 Deficiency
title_sort	proof-of-concept gene editing for the murine model of inducible arginase-1 deficiency
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/eae6a3ed458b46789cceae268bf9e7a9
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Proof-of-Concept Gene Editing for the Murine Model of Inducible Arginase-1 Deficiency

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