Figure 1.

Mouse Model for RPS-Deficient DBA and SIN Lentiviral Vectors for DBA Gene Therapy

Transgenic mice containing a doxycycline-regulatable Rps19-targeting shRNA allow an inducible and graded downregulation of Rps19. (A) Overview of modified loci. Black arrowheads indicate TSSs. (B) Breeding strategy to adjust the level of Rps19 downregulation. (C) EFS-RPS19 vector, codon-optimized human RPS19 cDNA was constructed under the control of the human elongation factor 1α short (EFS) promoter and inserted into a lentiviral vector. Following the RPS19 cDNA, an internal ribosomal entry site (IRES), a GFP sequence, and improved post-transcriptional regulatory element (Pre*) were inserted. EFS-Spacer vector, in which the RPS19 cDNA was replaced with an equally long non-coding spacer sequence, was used as a control. The LCR-EFS-RPS19 vector, where in locus control region of β-globin gene was inserted before the EFS promoter. LTR, long terminal repeat; pA, polyadenylation signal; PPT, polypurine tract; RRE, Rev response element; SA, splice acceptor; Wt, wild-type.