From siRNA to shRNA
Anchor the siRNA sequence ( 19nt ) on your target gene .
Extract 2nt of upstream + 19nt siRNA + 1nt of downstream , which is totally 22nt of nucleotides .
Antisense shRNA : Insert the reverse complement of the 22nt into shRNA backbone .
Sense shRNA : Insert the 22nt ( from step 2 ) into shRNA backbone .
Replace the 1st nt with other nucleotide ( replace A with G and vice versa ; replace C with T and vice versa ) .
Now your shRNA designing job is done .
You see , how easy it is !
Synthesis the shRNA nucleotides and insert them into pTRIPZ vector .
You may also translocate the shRNA from pTRIPZ vector into pGIPZ vector .
