I wonder if I mutate several base pairs of single strand DNA oligo in PAM and upstream from the Cas9 cleavage site at the same time.
Here is the original sequence and the desired changes. TGG is PAM sequence.
_ AGGTGCGC GTGCCAGTC GCG_ ** _TGG _ (original)**
-> AGGTGCGC AAA CCAGTC GCG AAA (desired change)
These Cas9 cleavage preventing mutations are the point mutations that I want to make and I do not need no other changes such as insertion or deletion.
Please let me know if this strategy works.