Optimized CRISPR-RfxCas13d system for RNA targeting in zebrafish embryos
Luis Hernández-Huertas, Gopal Kushawah, Alejandro Díaz‐Moscoso, Laura Tomás‐Gallardo, Ismael Moreno-Sánchez, Gabriel da Silva Pescador, Ariel Bazzini, Miguel A. Moreno-Mateos
Abstract
CRISPR-Cas systems have been used to induce DNA mutagenesis for gene function discovery. However, the development of tools to eliminate RNAs provides complementary and unique approaches to disrupt gene expression. Here, we present a workflow to perform specific, efficient, and cost-effective mRNA knockdown in zebrafish embryos using our in vivo optimized CRISPR-RfxCas13d (CasRx) system. Although the described protocol focuses on mRNA knockdown in zebrafish embryos, it can also be applied to other vertebrates. For complete details on the use and execution of this protocol, please refer to Kushawah et al. (2020).