For my internship I have to develop crispr-cas9 guides for certain genes. One of the things I have to validate in silico, is whether or not the crRNA has the ability to bind the tracRNA sequence when cloned into E. Coli. So far, any guide creating software I've found does not take this into account and I cannot seem to find any information about this. I personally think it's not needed to check this, but just to be sure I'd ask here if that would be needed.
Kind regards,
Chris
Uwe Borgmeyer
You are right. There is no need to validate binding of crRNA to tracRNA. The critical step is the efficiency and specificity of directing Cas9 to the desired location.
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