Design and Microinjection of Morpholino Antisense Oligonucleotides and mRNA into Zebrafish Embryos to Elucidate Specific Gene Function in Heart DevelopmentZain Zaki Zakaria 1,2, Shahram Eisa-Beygi 3, Fatiha M. Benslimane 1, Ramani Ramchandran 4, Huseyin C. Yalcin 1
1Biomedical Research Center, Qatar University, Doha, Qatar, 2Department of Biological and Environmental Sciences, College of Arts and Science, Qatar University, Doha, Qatar, 3Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, 4Department of Pediatrics, Medical College of Wisconsin
The present protocol describes designing, preparing, and microinjecting a translational-blocking morpholino against a representative cardiac gene; Heart And Neural Crest Derivatives Expressed2 (hand2) into the yolk of newly fertilized zebrafish embryos to knock down gene function. It also shows a transient rescue of these "morphants" by co-injection of mRNA encoding this gene product.
