Generation of a MYH6 (c.4034T > C) mutant human embryonic stem cell line via CRISPR base editing

Area of Science:

• Cardiovascular Genetics
• Stem Cell Biology
• Gene Editing Technologies

Background:

• The MYH6 gene is crucial for α-myosin heavy chain in the adult human heart.
• A specific MYH6 mutation (c.4034T>C, p.Leu1345Pro) is associated with hypertrophic cardiomyopathy (HCM), but its causal link remains uncertain.
• Establishing the unambiguous role of this mutation in HCM pathogenesis is critical for understanding the disease.

Purpose of the Study:

• To generate a human embryonic stem cell line carrying the MYH6 c.4034T>C mutation.
• To utilize CRISPR adenine base editing for precise genetic modification.
• To create a cellular model for studying the functional impact of the MYH6 mutation in HCM.

Main Methods:

• CRISPR adenine base editing was employed to introduce the MYH6 c.4034T>C mutation into human embryonic stem cells.
• The generated cell line (WAe009-A-1D) was characterized for its stem cell properties.
• In vivo differentiation potential was assessed to confirm pluripotency.

Main Results:

• A stable human embryonic stem cell line (WAe009-A-1D) harboring the MYH6 c.4034T>C mutation was successfully generated.
• The mutant cell line retained normal morphology, pluripotency, and karyotype.
• The cells demonstrated the capacity for in vivo differentiation into all three germ layers.

Conclusions:

• The developed MYH6 mutant human embryonic stem cell line provides a valuable tool for investigating the role of this specific mutation in hypertrophic cardiomyopathy.
• This cellular model allows for detailed mechanistic studies of MYH6-associated cardiac disease.
• The successful generation and characterization of this cell line advance the study of genetic cardiomyopathies.