It seems only mRNA or protein was used for cas9 injection in zebrafish. Is there anyone try plasmid cas9 DNA for zebrafish? what is the efficiency? Any good promoter for fish embryo? Is Nos1 good?
Yes, it is possible to use plasmid DNA encoding the Cas9 nuclease for gene editing in zebrafish. In fact, plasmid DNA has been widely used as a delivery vehicle for Cas9 in zebrafish and other model organisms, as it is relatively easy to produce and can be delivered using a variety of methods such as microinjection, electroporation, or hydrodynamic injection.
The efficiency of gene editing using plasmid Cas9 DNA in zebrafish may vary depending on a number of factors, including the specific target site, the method of delivery, and the presence of other genetic or environmental factors that may affect the efficiency of the editing process. In general, the efficiency of gene editing using Cas9 in zebrafish is relatively high, with reported rates of up to 70-80% for some target sites.
As for promoters, the choice of promoter can be important for driving the expression of Cas9 in zebrafish embryos. The Nos1 promoter has been used successfully to drive the expression of Cas9 in zebrafish embryos, and has been shown to be effective in a number of studies. Other promoters that have been used successfully in zebrafish include the hsp70 promoter, the zic1 promoter, and the cmv promoter. Ultimately, the choice of promoter will depend on the specific goals of the study and the desired pattern of gene expression in the embryos.
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