Now, Im working on using CRISPR tool to knock out TRPM2 gene in zebrafish. Is there any easy way to identify which is homo fish and which is hetero fish?
Hello Nam Ha.
There are over 10 family members in TRPM genes. And its properties seem to be transient. Hard to detect any phenotypic differences at early embryonic development. I do not know what tissues you are specifically looking for. However, before doing molecular works to screen, it may be detectable looking at the repsponses to the exposure to drugs (such as doxorubicin in both presence and absence of low CaCl2 sol, followed by DCF-DA staining to see ROS accumulation in specifically embryonic brain and heart regions.
Best wishes.
Your animals will likely be chimeras, not true knockouts (in the first generation). The T7 endonuclease assay (after PCR amplification of your region of interest) is probably the easiest way to know if the genome has been affected - if it works - it can be a challenging assay. You can also just sequence the PCR product, the sequence will likely just be a mess at the target site if the CRISPR is working.
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