How do I create forward and reverse gRNAs for dCas9 CRISPR?

21 August 2023 0 9K Report

Hello everyone! I am reading a protocol for CRISPR using a dCas9, and I came across the following passage in the gRNA section:

A. Design sgRNA oligos

Fwd (all): ACCGNNNNNNNNNNNNNNNNNNN

Rev (Sp): AAACNNNNNNNNNNNNNNNNNNN

PAM Choice:

Sp dCas9: NGG

How do you design forward and reverse gRNAs? I was under the impression that you only needed to create 1 gRNA. I am very confused!

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