I have generated a CRISPR/Cas9 knock-out zebrafish of my desired gene (referred to abc-/-) and an overexpression line of abc by using Tol2 transposon (referred to Tg(hsp7:abc)). All the phenotype characterisation and mechanism studies were done. However, one of the reviewers suggested that the loss of function of the abc-/- should be restored by overexpression of Tg(hsp7:abc). However, crossing the two lines together demand another 6 months and sampling injecting mRNA of abc to embryos is not suitable since we need to observe the behaviour phenotype at 7dpf and the injected mRNA will decay at that time.
Although I agree with the reviewer that the rescue experiment is necessary. But I think the overexpression lines have already shown the opposite phenotypes and not all the studies perform the rescue experiment. So I want to know if there are some other ways to respond to the reviewer.
Zexu Chen In this case, politely reply to the reviewer that the rescue experiment will take 6 months. Mention why you think your data is valid even without the rescue experiment, just like you did here, and in addition, explain if this experiment is the main experiment of the article or not. If not, mention it. Then, revise your text to use less harsh language, i.e. write in Discussion that "one of the potential drawbacks of our study is that no rescue experiment was performed". Then (in the answer to the reviewer) write "we have modified our manuscript to highlight the fact that the rescue experiment was not performed. Write then, "However, there are several papers where it was not performed (cite said papers), and conclude to the reviewer that "due to the time constraints and budget we ask the reviewer to consider modification of the article's text as an appropriate response to this problem".
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