Construction of gRNA Using CRISPR Gene Editing in Transgenic Danio Rio

Abstract

The CRISPR system can be used to perform site-directed mutagenesis in transgenic zebrafish lines while co-injecting a donor plasmid, short guided RNA (sgRNA), and Cas9 mRNA. In this study, the CRISPR system will be used to genetically edit DNA sequences of harpy mutants by targeting the hatching gland promoter (hgg) responsible for the early arrest mutation. Through molecular cloning techniques, polymerase chain reactions (PCR), and sequencing events, cleavage of the donor plasmid and the integration of donor DNA for one out of two vector clones successfully took place.