Telomerase is essential for cardiac differentiation and sustained metabolism of human cardiomyocytes II

In this study, we employed an inducible CRISPRi human induced pluripotent stem cell (hiPSC) line to silence TERT expression enabling the generation of hiPSCs and hiPSC-derived cardiomyocytes with long and short telomeres. Reduced telomerase activity and shorter telomere lengths of hiPSCs strongly hampered their differentiation potential towards cardiomyocytes Overall design: The bulk RNA seq was performed on CRISPRi TERT hiPSC line with 4 different conditions in a doxycycline inducible CRISPRi TERT hiPSC line. The telomerase activity and telomere length were altered at the hiPSC stage to give rise to - long telomeres (C2 group, no doxycycline), intermediate telomeres (D2 group- 2 passages with doxycycline) and short telomeres (D5 group-5 passages with doxycycline) and reversed (R2 group-2 passages without doxycycline from the D5 group).

本研究采用诱导型CRISPR干扰（CRISPRi）人类诱导多能干细胞（human induced pluripotent stem cell, hiPSC）细胞系，通过沉默端粒酶逆转录酶（telomerase reverse transcriptase, TERT）的表达，成功获得了具有长、短端粒的hiPSC及其分化而来的心肌细胞。研究发现，hiPSC的端粒酶活性降低与端粒长度缩短，会显著抑制其向心肌细胞分化的潜能。

实验整体设计：
本研究针对该多西环素诱导型CRISPRi TERT hiPSC细胞系的4种不同条件样本开展批量RNA测序（bulk RNA-seq）。我们在hiPSC阶段调控端粒酶活性与端粒长度，得到四组样本：长端粒组（C2组，不添加多西环素）、中端粒组（D2组，经2次传代并添加多西环素）、短端粒组（D5组，经5次传代并添加多西环素）以及恢复组（R2组，取自D5组后经2次传代且不添加多西环素）。