sgRNA design

Question

[elisa.qinxue]

Hello,

I'm used ChopChop to design sgRNA used for Crispr gene editing. As you see on the image, why the sgRNA were marked in two directions when the sequences are all in the forward direction? What's the use of the direction?

Thank you in advance!

Answer 1

[ATpoint]

gRNAs are single-stranded and by this they have a strand-specificity (depends where the PAM is) and that gives them an orientation.

[elisa.qinxue]

So it would be OK no matter where I would like to cut if I use appropriate sgRNA, like below.