Fig. 1.

Schematic drawings of shRNA constructs for in vitro/in vivo studies. a For in vitro studies, the short hairpin RNAs were cloned in pBluescript vector under the control of the U6 promoter (U6). b Sequences of the hairpins tested for their downregulation efficiency via Western blot, after electroporation of the shRNA vector constructs into ES cells. Sequences of the hairpin-loops are underlined. c Schematic representation of the recombinant AAV vector genome AAV-CMV-Venus-U6shRNA. The two expression cassettes for the reporter Venus and the shRNA are separated by a 1.0 kb stuffer element and the genome is flanked by two 141 bp inverted terminal repeats. ITR inverted terminal repeats; CMV cytomegalovirus promoter; intron: β-globin intron; Venus: yellow fluorescent protein Venus; pA: human growth hormone polyA signal; stuffer: stuffer element; U6: U6 promoter; LacZsh: control hairpin; Erksh4: hairpin against Erk2