Hello, I'm trying to design an array of 4 crRNAs for use with cpf1/cas12a both the As and Lb versions. I've found the scaffold and PAM sequence from digging through a few papers, but I can't find a resource that explicitly states what sequence to use between the individual crRNAs so they will be cleaved by cpf1/cas12a. Does anyone know what this sequence would be or where to find it? Or even better, a publication that explicitly lists the sequence of all components (scaffold, PAM, and processing site). Thank you!
Cas12a can process its own crRNA so in general, it does not need any specific sequence between two gRNA arrays.
But there is a report about boosting its processing using tRNA-crRNA arrays. https://www.pnas.org/content/112/11/3570
I hope you may find sequences of PAM, scaffold, and others you need in the supplementary information. But if you still need more guidance please contact me. I will be happy to help you.
Hi...they're processed at the DR (direct repeats)...this methods paper (actually the supplemental that goes with it) has what you're looking for (mainly for the As version)
Article Multiplexed genome engineering by Cas12a and CRISPR arrays e...
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