Stimulation of Cardiomyogenisis by CRISPR Cas9 approach

Myocardial infarction (MI) causes substantial damage to cardiovascular tissues due to hypoxia-induced apoptosis. Heart failure is a leading cause of death following ischemia-reperfusion injury. To avoid heart failure after MI, cardiomyocyte proliferation must be induced. Unfortunately, the adult heart has limited regenerative capacities primarily due to a signaling cascade known as Hippo pathway, activated by TAO kinases 1, 2, and 3 (TAOK 1-3 ). While Hippo pathway activation is crucial for preventing cancer development, it also inhibits cellular regeneration. Deactivation of the Hippo pathway is crucial for the proliferation of cardiomyocytes. An active Hippo pathway will phosphorylate the YAP/TAZ co-factors, hindering them from stimulating cellular regeneration. If the Hippo pathway is deactivated, the YAP/TAZ co-factors are able to facilitate the reentry of cardiomyocytes into the cell cycle by interacting with TEAD transcription factor to express downstream target genes. Here we suggest deactivating Hippo pathway through the conditional knockout (KO) of the TAOK 1-3 from human induced pluripotent stem cell cardiomyocytes (hiPSC-CMs) by CRISPR Cas-9 to induce proliferation of cardiomyocytes. HiPSC-CMs will be matured to resemble adult cardiomyocyte physiology. Elimination of TAOK 1-3 will deactivate Hippo pathway and prevent phosphorylation of YAP/TAZ co-factors which will stimulate cardiomyocyte proliferation. Once the cardiomyocytes have been transformed, the cells should undergo replication after the event of an ischemia-reperfusion injury. Cardiomyogenesis will be assessed in the control and KO cells. By inhibiting expression of  TAOK 1-3, cardiomyocyte proliferation should occur, resulting in the regeneration of cardiovascular tissues and preventing heart failure after MI.