Hello,

si it seems you just need to induce a KO in each of your specific genes, right?

So you juste have to design some sgRNA (you can use this webtool: http://crispor.tefor.net/crispor.py)

so add your genomic sequence for the gene you want to KO (first exon usually), select the Cas9 protein you will be using (typically S. pyogenes, that recognizes a "NGG" motif). Then you just have to select the different sgRNA you will be using, with the activity and specificity scores.

then, when you click on the sgRNA you choose, a new window appears, with the sequences of the oligos you have to order to clone them into a vector (you can order it from addgene), and then express them into the cells, with Cas9 vector (also available from addgene).

Then you will have to clone the cells, to isolate a mutated cell, and let it grow. (you should perform before a T7 assay, to know your mutation rate, to know how many cells you have to clone, to isolate enough mutated clones)

If i'm not clear and you need more infos, feel free to ask me, i work in a genome editing lab.

Yes, you are right. And thank you very much for your answer!!

I'd suggest http://www.origene.com/crispr-cas9/

The kit has all the essentials you need:

Complete kit for gene knockout using CRISPR

Two guide RNA vectors in for efficient cleavage

Knockin GFP-Puro for selection

pCas-Guide-scramble included as a negative control

Thank you Tin-Lap and Yujiro!!  Those are nice option.