Abstract
ObjectivesTo investigate the effect of endogenous Cas9 on genome editing efficiency in transgenic zebrafish.ResultsHere we have constructed a transgenic zebrafish strain that can be screened by pigment deficiency. Compared with the traditional CRISPR injection method, the transgenic zebrafish can improve the efficiency of genome editing significantly. At the same time, we first observed that the phenotype of vertebral malformation in early embryonic development of zebrafish after ZFERV knockout.ConclusionsThe transgenic zebrafish with expressed Cas9, is more efficient in genome editing. And the results of ZFERV knockout indicated that ERV may affect the vertebral development by Notch1/Delta D signal pathway.
Highlights
Zebrafish are a classical animal model for developmental biology and biomedical studies; several important questions in vertebrate development have been addressed using this model
The transgenic zebrafish with expressed Cas9, is more efficient in genome editing
The results of zebrafish ERV (ZFERV) knockout indicated that Endogenous retroviruses (ERVs) may affect the vertebral development by Notch1/ Delta D signal pathway
Summary
Zebrafish are a classical animal model for developmental biology and biomedical studies; several important questions in vertebrate development have been addressed using this model. Large-scale screening in the 1990s for zebrafish mutants generated via random mutagenesis resulted in zebrafish becoming one of the most frequently used animal models. Newly developed genome-editing tools, including ZFNs, TALENs, and CRISPR/Cas, provide a feasible approach for the generation of zebrafish with precise genomic modifications. These zebrafish have significantly promoted the understanding of genes with unknown functions, especially those involved in embryonic development (Jao et al 2013). Establishing a zebrafish homozygous line with a genomic modification from the mosaic F0 generation of zebrafish is time consuming, requiring breeding and genotyping of the F1 and F2 generations (Varshney et al 2015).
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