I am looking for designing crRNA against the putative G-quadruplex forming motifs. Taking account of the criteria for a good crRNA, web-based design gives crRNA sequences that don't overlap the sequence of my interest. On the contrary, manual design compromises the quality of the crRNA with lots of off-targets in the genome. However, this is very difficult in designing any primer against the sequence having tandem guanine repeats.
Therefore, I have planned to design two crRNAs in (+) and (-) strands respective having 84 bases in between and want to ensure the deletion of the 84 bases in between by CRISPR-Cas9 method. My design of crRNA is provided in the attachment. Please help me in in this regard.