Insertional vs Targeted Mutagenesis in the Development of Zebrafish as an In Vivo Model for Cardiomyopathy

Heart failure is a global economic burden and can be caused by cardiomyopathy, a disease of the myocardium. Mutations in genes encoding sarcomere proteins have been known to cause cardiomyopathy. One of these sarcomere proteins is α-cardiac actin (ACTC), which is needed for proper contraction of the heart. To better understand how mutations in the ACTC protein cause cardiomyopathy, an in vivo model like zebrafish can be used to understand the molecular mechanism that occurs. Using transposons (insertional) or Clustered Regularly Interspaced Palindromic Repeats (CRISPRs) (targeted), zebrafish can be engineered to carry these mutations and the impact can be studied. The transposon system proved to be unstable as the inserted transposon underwent transcriptional repression. To use the CRISPR system, identifying which zfactc gene to target is necessary. Performing WMISH and RT-qPCR, zfacta1b seems to be the best candidate for targeted mutagenesis due to its early expression in the heart.