Multi-gene duplication removal in an engineered human cellular MECP2 duplication syndrome model with an IRAK1-MECP2 duplication

Recent progress in genome editing technologies has catalyzed the generation of sophisticated cell models; however, the precise modeling of copy-number variation (CNV) diseases remains a significant challenge despite their substantial prevalence in the human population. To overcome this barrier, we h...

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Main Authors: Samar Z. Rizvi, Wing Suen Chan, Eleonora Maino, Sydney Steiman, Georgiana Forguson, Maya Klepfish, Ronald D. Cohn, Evgueni A. Ivakine
Format: Article
Language:English
Published: Elsevier 2024-12-01
Series:Molecular Therapy: Nucleic Acids
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Online Access:http://www.sciencedirect.com/science/article/pii/S2162253124002439
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author Samar Z. Rizvi
Wing Suen Chan
Eleonora Maino
Sydney Steiman
Georgiana Forguson
Maya Klepfish
Ronald D. Cohn
Evgueni A. Ivakine
author_facet Samar Z. Rizvi
Wing Suen Chan
Eleonora Maino
Sydney Steiman
Georgiana Forguson
Maya Klepfish
Ronald D. Cohn
Evgueni A. Ivakine
author_sort Samar Z. Rizvi
collection DOAJ
description Recent progress in genome editing technologies has catalyzed the generation of sophisticated cell models; however, the precise modeling of copy-number variation (CNV) diseases remains a significant challenge despite their substantial prevalence in the human population. To overcome this barrier, we have explored the utility of HAP1 cells for the accurate modeling of disease genomes with large structural variants. As an example, this study details the strategy to generate a novel cell line that serves as a model for the neurological disorder methyl CpG binding protein 2 (MECP2) duplication syndrome (MDS), featuring the critical duplication of both the MECP2 and IRAK1 genes. This model faithfully recapitulates MDS genomic rearrangement, allowing for the mechanistic study of gene overexpression and the development of therapeutic interventions. Employing a single-guide RNA (gRNA) CRISPR-Cas9 strategy, we successfully excised the duplicated genomic segment, notably halving both MECP2 and IRAK1 expression levels. The evidence establishes our model as a crucial tool for research into MDS. Furthermore, the outlined workflow is readily adaptable to model other CNV disorders and subsequently test genomic and pharmacological interventions.
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spelling doaj-art-16631687f3624b21a3da29b4dc879eca2025-08-20T01:54:15ZengElsevierMolecular Therapy: Nucleic Acids2162-25312024-12-0135410235610.1016/j.omtn.2024.102356Multi-gene duplication removal in an engineered human cellular MECP2 duplication syndrome model with an IRAK1-MECP2 duplicationSamar Z. Rizvi0Wing Suen Chan1Eleonora Maino2Sydney Steiman3Georgiana Forguson4Maya Klepfish5Ronald D. Cohn6Evgueni A. Ivakine7Program in Genetics and Genome Biology, The Hospital for Sick Children Research Institute, Toronto, ON M5G 0A4, Canada; Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, CanadaProgram in Genetics and Genome Biology, The Hospital for Sick Children Research Institute, Toronto, ON M5G 0A4, CanadaBiozentrum, The Center for Molecular Life Sciences, University of Basel, Basel 4056, SwitzerlandProgram in Genetics and Genome Biology, The Hospital for Sick Children Research Institute, Toronto, ON M5G 0A4, Canada; Department of Physiology, University of Toronto, Toronto, ON M5S 1A8, CanadaProgram in Genetics and Genome Biology, The Hospital for Sick Children Research Institute, Toronto, ON M5G 0A4, CanadaProgram in Genetics and Genome Biology, The Hospital for Sick Children Research Institute, Toronto, ON M5G 0A4, CanadaProgram in Genetics and Genome Biology, The Hospital for Sick Children Research Institute, Toronto, ON M5G 0A4, Canada; Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada; Institute of Medical Science, University of Toronto, Toronto, ON M5S 1A8, Canada; Department of Pediatrics, The Hospital for Sick Children, Toronto, ON M5G 1E8, CanadaProgram in Genetics and Genome Biology, The Hospital for Sick Children Research Institute, Toronto, ON M5G 0A4, Canada; Department of Molecular Genetics, University of Toronto, Toronto, ON M5S 1A8, Canada; Corresponding author: Evgueni A. Ivakine, Program in Genetics and Genome Biology, The Hospital for Sick Children Research Institute, Toronto, ON M5G 0A4, Canada.Recent progress in genome editing technologies has catalyzed the generation of sophisticated cell models; however, the precise modeling of copy-number variation (CNV) diseases remains a significant challenge despite their substantial prevalence in the human population. To overcome this barrier, we have explored the utility of HAP1 cells for the accurate modeling of disease genomes with large structural variants. As an example, this study details the strategy to generate a novel cell line that serves as a model for the neurological disorder methyl CpG binding protein 2 (MECP2) duplication syndrome (MDS), featuring the critical duplication of both the MECP2 and IRAK1 genes. This model faithfully recapitulates MDS genomic rearrangement, allowing for the mechanistic study of gene overexpression and the development of therapeutic interventions. Employing a single-guide RNA (gRNA) CRISPR-Cas9 strategy, we successfully excised the duplicated genomic segment, notably halving both MECP2 and IRAK1 expression levels. The evidence establishes our model as a crucial tool for research into MDS. Furthermore, the outlined workflow is readily adaptable to model other CNV disorders and subsequently test genomic and pharmacological interventions.http://www.sciencedirect.com/science/article/pii/S2162253124002439MT: RNA/DNA EditingMECP2 duplication syndromeIRAK1disease modelingHAP1CRISPR-Cas9
spellingShingle Samar Z. Rizvi
Wing Suen Chan
Eleonora Maino
Sydney Steiman
Georgiana Forguson
Maya Klepfish
Ronald D. Cohn
Evgueni A. Ivakine
Multi-gene duplication removal in an engineered human cellular MECP2 duplication syndrome model with an IRAK1-MECP2 duplication
Molecular Therapy: Nucleic Acids
MT: RNA/DNA Editing
MECP2 duplication syndrome
IRAK1
disease modeling
HAP1
CRISPR-Cas9
title Multi-gene duplication removal in an engineered human cellular MECP2 duplication syndrome model with an IRAK1-MECP2 duplication
title_full Multi-gene duplication removal in an engineered human cellular MECP2 duplication syndrome model with an IRAK1-MECP2 duplication
title_fullStr Multi-gene duplication removal in an engineered human cellular MECP2 duplication syndrome model with an IRAK1-MECP2 duplication
title_full_unstemmed Multi-gene duplication removal in an engineered human cellular MECP2 duplication syndrome model with an IRAK1-MECP2 duplication
title_short Multi-gene duplication removal in an engineered human cellular MECP2 duplication syndrome model with an IRAK1-MECP2 duplication
title_sort multi gene duplication removal in an engineered human cellular mecp2 duplication syndrome model with an irak1 mecp2 duplication
topic MT: RNA/DNA Editing
MECP2 duplication syndrome
IRAK1
disease modeling
HAP1
CRISPR-Cas9
url http://www.sciencedirect.com/science/article/pii/S2162253124002439
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