I knocked out a specific gene in zebrafish for study by the CRISPR cas9 system. In the F0 founder fish, there are about 80% of fish with mutations in a certain gene. For one fish with mutations in F0, there are lots of mutations(different types of insertions or deletions) in the germ cell. I wondered how these different types of mutation formed/developed? Can somebody give me some explanations? Thanks so much!!
CRISPR is not a perfect system, it's just generally easier to use than other genetic engineering systems (e.g. TALEN, ZFNs) that came before it. Off-target cleavage is a common concern when using CRISPR, and you should check the specificity of your sgRNA using something like https://cctop.cos.uni-heidelberg.de:8043.
I agree with John and you should check your HDR- fragment if you are giving any.
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