Does directionality matter in this case? Will the said sgRNA still bind properly to my region of interest and recruit cas9? Thank you so much for your thoughts!
Margs S Brennan
If I understand your question correctly, yes it will still bind and recruit Cas9. In other words you can design your sgRNA to bind either the coding OR non-coding strand, as long as the PAM sequence is still at the 3'end. It may be useful to keep in mind that Cas9 will cleave your target DNA a few nucleotides upstream from the PAM site.
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