While designing theguide RNA for CRISPR we prefer to use two guide RNA for greater knockout efficiency, So can we design the two guide RNA one from sense and other from antisense strand?
If your intention is knockout by means of a DSB (Double Strand Break), it is the same whether your guide RNAs are on the sense or antisense strand. They just have to be within a few bp of each other so as not to interfere with each other.
Only in the case of using a Cas9 nickase you have to be aware of which strand do you need for the design