This vector uses a bidirectional U6 promoter to express two small hairpin RNA sequences that can silence two genes simultaneously. Two short dsDNAs that encode two shRNAs to target the same gene or two different genes are ligated into two pairs of restriction sites (Mlu I/Spe I and Bgl II/Not I) at each side of the promoter. The dsDNAs are constructed by annealing complementary synthetic oligonucleotides. Each oligonucleotide is designed with a 4-base overhang at its 5’-end that complements the corresponding restriction site. The other features of the plasmid facilitate its rapid conversion to an adenovirus vector using the SpeAd procedure (see Kit #K2210).

---