Figure 1
From: Design and cloning strategies for constructing shRNA expression vectors
Design strategies for creating short hairpin RNA (shRNA) template inserts. (a) Expressed shRNA is transcribed as a ssRNA molecule that folds onto itself forming a stem-loop structure. (b) Annealed complementary oligos can be used to create a synthetic DNA duplex (74 % of studies) for cloning. (c) Similar inserts for cloning can be made as complete promoter-shRNA cassettes by PCR using a hairpin containing primer (22 % of studies). (d) Alternatively, extension of a generic primer bound to a unique 'template' oligo can create a synthetic insert, akin to the annealed complementary oligo strategy (4 % of studies).