| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | CRISPR/Cas9-mediated gene editing was performed as described before50–53. Briefy, ATG knock-out of troponin T was performed by oligonucleotide annealing the sgRNA sequences (5′- GACCATGTCTGACATAGAAG-3′, 5′-CTTCTATGTCAGACATGGTC-3′), subcloning into the pSpCas9(BB)-2A-Puro plasmid and subsequent iPSC transfection and single clone isolation. pcDNA 3.1 plasmids carrying the full-length TNNT2-C-terminal-DYK sequence were purchased from GenScript. | Get A Quote |
The sarcomeric troponin-tropomyosin complex is a critical mediator of excitation-contraction coupling, sarcomeric stability and force generation. We previously reported that induced pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) from patients with a dilated cardiomyopathy (DCM) mutation, troponin T (TnT)-R173W, display sarcomere protein misalignment and impaired contractility. Yet it is not known how TnT mutation causes dysfunction of sarcomere microdomains and how these events contribute to misalignment of sarcomeric proteins in presence of DCM TnT-R173W. Using a human iPSC-CM model combined with CRISPR/Cas9-engineered isogenic controls, we uncovered that TnT-R173W destabilizes molecular interactions ... More
