Editing Fish Genome With CRISPR

The advent of CRISPR genome editing has revolutionized genetic analysis. We have adapted CRISPR for targeted gene disruption in zebrafish. We show that delivery of in vivo synthesized single guide RNA (sgRNA) and mRNA for a codon optimized Cas9 by microinjection results in efficient biallelic gene inactivation in somatic cells, allowing rapid functional assessment in the injected fish.  In addition, several genes can be disrupted simultaneously by injecting multiples sgRNAs. Furthermore, we show that transgenic expression of sgRNA and Cas9 is sufficient to cause biallelic mutagenesis, and tissue specific and/or inducible expression of Cas9 affords conditional mutagenesis. Finally, I will discuss our work on generating point mutations and targeted integrations using CRISPR.