Cre-Controlled CRISPR mutagenesis provides fast and easy conditional gene inactivation in zebrafish

Abstract Conditional gene inactivation is a powerful tool to determine function when constitutive mutations result in detrimental effects. The most commonly used technique achieve conditional employs the Cre/loxP system and its ability delete DNA sequences flanked by two loxP sites. However, targeting with sites time labor consuming. Here, we show Cre-Controlled CRISPR (3C) mutagenesis circumvent these issues. 3C relies on gRNA Cre-dependent Cas9-GFP expression from same transgene. Exogenous or transgenic supply of Cre results subsequent interest. recombined cells become fluorescently visible enabling their isolation subjection various omics techniques. Hence, provides valuable alternative production loxP-flanked alleles. It might even enable multiple genes simultaneously should be applicable other model organisms amenable single integration transgenesis.