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CRISPR-based knockout and base editing confirm the role of MYRF in heart development and congenital heart disease

High-throughput DNA sequencing studies increasingly associate DNA variants with congenital heart disease (CHD). However, functional modeling is a crucial prerequisite for translating genomic data into clinical care. We used CRISPR-Cas9-mediated targeting of 12 candidate genes in the vertebrate model...

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Main Authors: Doering, Lino (Author) , Cornean, Alex (Author) , Thumberger, Thomas (Author) , Benjaminsen, Jørgen (Author) , Wittbrodt, Beate (Author) , Kellner, Tanja (Author) , Hammouda, Omar T. (Author) , Gorenflo, Matthias (Author) , Wittbrodt, Joachim (Author) , Gierten, Jakob (Author)
Format: Article (Journal)
Language:English
Published: August 2023
In: Disease models & mechanisms
Year: 2023, Volume: 16, Issue: 8, Pages: 1-14
ISSN:1754-8411
DOI:10.1242/dmm.049811
Online Access:Verlag, kostenfrei, Volltext: https://doi.org/10.1242/dmm.049811
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Author Notes:Lino Doering, Alex Cornean, Thomas Thumberger, Joergen Benjaminsen, Beate Wittbrodt, Tanja Kellner, Omar T. Hammouda, Matthias Gorenflo, Joachim Wittbrodt and Jakob Gierten
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Summary:High-throughput DNA sequencing studies increasingly associate DNA variants with congenital heart disease (CHD). However, functional modeling is a crucial prerequisite for translating genomic data into clinical care. We used CRISPR-Cas9-mediated targeting of 12 candidate genes in the vertebrate model medaka (Oryzias latipes), five of which displayed a novel cardiovascular phenotype spectrum in F0 (crispants): mapre2, smg7, cdc42bpab, ankrd11 and myrf, encoding a transcription factor recently linked to cardiac-urogenital syndrome. Our myrf mutant line showed particularly prominent embryonic cardiac defects recapitulating phenotypes of pediatric patients, including hypoplastic ventricle. Mimicking human mutations, we edited three sites to generate specific myrf single-nucleotide variants via cytosine and adenine base editors. The Glu749Lys missense mutation in the conserved intramolecular chaperon autocleavage domain fully recapitulated the characteristic myrf mutant phenotype with high penetrance, underlining the crucial function of this protein domain. The efficiency and scalability of base editing to model specific point mutations accelerate gene validation studies and the generation of human-relevant disease models.
Item Description:Online veröffentlicht: 16. August 2023
Gesehen am 23.11.2023
Physical Description:Online Resource
ISSN:1754-8411
DOI:10.1242/dmm.049811

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