An efficient dissociation protocol for generation of single cell suspension from zebrafish embryos and larvae

Abstract

Zebrafish (Danio rerio) has emerged as a powerful animal model to study developmental processes and human diseases. The introduction of CRISPR/Cas9 as a genome editing tool allowed the generation of genetic mutants with high-throughput (Varshney et al., 2015) and has opened the possibility to understand gene function not only during embryonic stages but also in larval stages. Therefore, there is an increasing need to optimize methods for embryo and larvae dissociation that allow the generation of single cell suspension for fluorescence-activated cell sorting (FACS), RNA extraction and single cell RNA-sequencing.

Keywords: Dissociation; Larvae dissociation to single cell suspension; Single cell suspension; Zebrafish; Zebrafish embryo.

Graphical abstract

Fig. 1

Visual representation of the 3-Step protocol for zebrafish embryos and larvae dissociation.