Fig. 1

Design of the rAAV9 mediated miR-29b transduction in vivo. A Schematic representation of miR-29b expression cassette. The expression cassette was comprised of miR-29b stem-loop sequence flanked by its native intron sequence, which preserves the putative hairpin structure and proper endogenous processing. A genomic fragment 800 bp containing the miR-29b precursor was PCR amplified from the mouse miR-29b precursor sequence and cloned into the self-complementary rAAV9. miR-29b under the control of cytomegalovirus and U6 promoter. rAAV9-GFP transduction in renal tissue was tested by GFP B western blot and C fluorescence in fresh frozen sections of kidney. In AAV-injected kidneys, there is significant EGFP fluorescence in tubule epithelial cells at 5 weeks. D Overall design of the study in vivo. Mouse were subjected to continuous AngII or PBS infusion. Two weeks later, when fibrosis was evident, the animals were randomly chosen to receive either recombinant adeno-associated vector type 9 carrying miR-29b (rAAV9-miR-29b) or control vector (rAAV9-random miR) at 5*10¹⁰ IVP per kidney via in situ injection