I want to start with some CRISPR experiments (with the nickase system) and I am wondering what sequences I should use for a non-target control?
I am probably overthinking this, but at the moment I really cannot come up with anything.. any suggestions?
Thanks in advance,
Sophie
Response to your question, for the sequence of the non target control you need to get known of the inverse of genetic programming for system identification. the simplest thing to target is by using least square method and for non target is to make it return back system identification graph back to zeros.
Article Genetic programming for system identification
When you design your CRISPR guide RNAs using the mit website, it generates a list of sequences that can be off target binding sites for your guides. I think you can select any of these off target regions to design your non-target control. Ideally if your CRISPR guides are specific to your target site, doing a PCR to amplify the predicted off target or non-target region should show you no in/dels. Does that answer your question? Please give me feedback, since I am new to CRISPR too!
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