In vivo genome editing at the albumin locus to treat methylmalonic acidemia

Methylmalonic acidemia (MMA) is a metabolic disorder most commonly caused by mutations in the methylmalonyl-CoA mutase (MMUT) gene. Although adeno-associated viral (AAV) gene therapy has been effective at correcting the disease phenotype in MMA mouse models, clinical translation may be impaired by l...

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Autores principales:	Jessica L. Schneller, Ciaran M. Lee, Leah E. Venturoni, Randy J. Chandler, Ang Li, Sangho Myung, Thomas J. Cradick, Ayrea E. Hurley, William R. Lagor, Gang Bao, Charles P. Venditti
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	adeno-associated virus albumin CRISPR-Cas9 metabolic disorders methylmalonic acidemia genome editing Genetics QH426-470 Cytology QH573-671
Acceso en línea:	https://doaj.org/article/40477017c9594449b2c47c26ce788c14
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spelling	oai:doaj.org-article:40477017c9594449b2c47c26ce788c142021-11-28T04:33:16ZIn vivo genome editing at the albumin locus to treat methylmalonic acidemia2329-050110.1016/j.omtm.2021.11.004https://doaj.org/article/40477017c9594449b2c47c26ce788c142021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2329050121001765https://doaj.org/toc/2329-0501Methylmalonic acidemia (MMA) is a metabolic disorder most commonly caused by mutations in the methylmalonyl-CoA mutase (MMUT) gene. Although adeno-associated viral (AAV) gene therapy has been effective at correcting the disease phenotype in MMA mouse models, clinical translation may be impaired by loss of episomal transgene expression and magnified by the need to treat patients early in life. To achieve permanent correction, we developed a dual AAV strategy to express a codon-optimized MMUT transgene from Alb and tested various CRISPR-Cas9 genome-editing vectors in newly developed knockin mouse models of MMA. For one target site in intron 1 of Alb, we designed rescue cassettes expressing MMUT behind a 2A-peptide or an internal ribosomal entry site sequence. A second guide RNA targeted the initiator codon, and the donor cassette encompassed the proximal albumin promoter in the 5′ homology arm. Although all editing approaches were therapeutic, targeting the start codon of albumin allowed the use of a donor cassette that also functioned as an episome and after homologous recombination, even without the expression of Cas9, as an integrant. Targeting the albumin locus using these strategies would be effective for other metabolic disorders where early treatment and permanent long-term correction are needed.Jessica L. SchnellerCiaran M. LeeLeah E. VenturoniRandy J. ChandlerAng LiSangho MyungThomas J. CradickAyrea E. HurleyWilliam R. LagorGang BaoCharles P. VendittiElsevierarticleadeno-associated virusalbuminCRISPR-Cas9metabolic disordersmethylmalonic acidemiagenome editingGeneticsQH426-470CytologyQH573-671ENMolecular Therapy: Methods & Clinical Development, Vol 23, Iss , Pp 619-632 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	adeno-associated virus albumin CRISPR-Cas9 metabolic disorders methylmalonic acidemia genome editing Genetics QH426-470 Cytology QH573-671
spellingShingle	adeno-associated virus albumin CRISPR-Cas9 metabolic disorders methylmalonic acidemia genome editing Genetics QH426-470 Cytology QH573-671 Jessica L. Schneller Ciaran M. Lee Leah E. Venturoni Randy J. Chandler Ang Li Sangho Myung Thomas J. Cradick Ayrea E. Hurley William R. Lagor Gang Bao Charles P. Venditti In vivo genome editing at the albumin locus to treat methylmalonic acidemia
description	Methylmalonic acidemia (MMA) is a metabolic disorder most commonly caused by mutations in the methylmalonyl-CoA mutase (MMUT) gene. Although adeno-associated viral (AAV) gene therapy has been effective at correcting the disease phenotype in MMA mouse models, clinical translation may be impaired by loss of episomal transgene expression and magnified by the need to treat patients early in life. To achieve permanent correction, we developed a dual AAV strategy to express a codon-optimized MMUT transgene from Alb and tested various CRISPR-Cas9 genome-editing vectors in newly developed knockin mouse models of MMA. For one target site in intron 1 of Alb, we designed rescue cassettes expressing MMUT behind a 2A-peptide or an internal ribosomal entry site sequence. A second guide RNA targeted the initiator codon, and the donor cassette encompassed the proximal albumin promoter in the 5′ homology arm. Although all editing approaches were therapeutic, targeting the start codon of albumin allowed the use of a donor cassette that also functioned as an episome and after homologous recombination, even without the expression of Cas9, as an integrant. Targeting the albumin locus using these strategies would be effective for other metabolic disorders where early treatment and permanent long-term correction are needed.
format	article
author	Jessica L. Schneller Ciaran M. Lee Leah E. Venturoni Randy J. Chandler Ang Li Sangho Myung Thomas J. Cradick Ayrea E. Hurley William R. Lagor Gang Bao Charles P. Venditti
author_facet	Jessica L. Schneller Ciaran M. Lee Leah E. Venturoni Randy J. Chandler Ang Li Sangho Myung Thomas J. Cradick Ayrea E. Hurley William R. Lagor Gang Bao Charles P. Venditti
author_sort	Jessica L. Schneller
title	In vivo genome editing at the albumin locus to treat methylmalonic acidemia
title_short	In vivo genome editing at the albumin locus to treat methylmalonic acidemia
title_full	In vivo genome editing at the albumin locus to treat methylmalonic acidemia
title_fullStr	In vivo genome editing at the albumin locus to treat methylmalonic acidemia
title_full_unstemmed	In vivo genome editing at the albumin locus to treat methylmalonic acidemia
title_sort	in vivo genome editing at the albumin locus to treat methylmalonic acidemia
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/40477017c9594449b2c47c26ce788c14
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In vivo genome editing at the albumin locus to treat methylmalonic acidemia

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