Retina-specific expression from the IRBP promoter in transgenic mice is conferred by 212 bp of the 5′-flanking region*

https://doi.org/10.1016/S0006-291X(05)81395-6Get rights and content

IRBP is a photoreceptor-specific glycoprotein that has been suggested as a retinoid carrier in the visual process. Previous research has shown that 1.3 kb of 5′-flanking sequence from the human IRBP gene is sufficient to promote photoreceptor-specific expression of reporter genes in transgenic mice. To define more narrowly the sequences that promote tissue-specific expression, chimeric constructs with shorter promoters were used to generate transgenic mice. The bacterial CAT gene was fused to fragments of 706 bp or 212 bp from the 5′ end of the human IRBP gene. Analysis of the three transgenic families bearing the 706 bp IRBP promoter revealed that CAT expression was confined to the neuro-retina and the pineal gland. Analysis of the four transgenic families bearing the 212 bp IRBP promoter revealed the same tissue-specific CAT expression in three families. These results establish that tissue-specific expression of IRBP can be regulated by a short 212 bp promoter which has been conserved between humans and mice.

References (18)

  • Carter-DawsonL. et al.

    Dev. Biol.

    (1986)
  • van VeenT. et al.

    Exp. Eye. Res.

    (1988)
  • LiouG.I. et al.

    J. Biol. Chem.

    (1990)
  • LiouG.I. et al.

    J. Biol. Chem.

    (1989)
  • DaleR.M.K. et al.

    Plasmid

    (1985)
  • ShawW.V.

    Methods Enzymol.

    (1975)
  • ChaderG.J.

    Invest. Ophthalmol. Vis. Sci.

    (1989)
  • LiouG.I. et al.
  • LiouG.I. et al.
There are more references available in the full text version of this article.

Cited by (37)

  • Triple Vectors Expand AAV Transfer Capacity in the Retina

    2018, Molecular Therapy
    Citation Excerpt :

    In order to test transduction efficiency mediated by triple AAV vectors, we generated a reporter protein by fusing the CDS of EGFP to that of Discosoma red fluorescent protein (DsRed) (herein referred to as ED) separated by a 13 amino acid spacer. A triple flag tag (3xflag) was added at the 3′ end of ED CDS (Figure 1A), which was placed under the control of (1) the ubiquitous cytomegalovirus (CMV) promoter, (2) the PR-specific human interphotoreceptor retinoid-binding protein (IRBP) promoter,41,42 or (3) the RPE-specific vitelliform macular dystrophy 2 (VMD2) promoter.43 The ED cassettes were either packed in a single AAV vector or split in three parts, each packed in a different AAV vector (Figure 1B and Materials and Methods) from here called ED-AAV 1, ED-AAV 2, ED-AAV 3.

  • Nanoparticles for retinal gene therapy

    2010, Progress in Retinal and Eye Research
    Citation Excerpt :

    The nanoparticles used for these studies were compacted with acetate as the lysine counterion forming rods, and they contained an expression cassette which had 1 of 3 promoters preceding the normal murine RDS cDNA (termed NMP) (Cai et al., 2009a,b). The first promoter derived from the interphotoreceptor retinoid binding protein (IRBP) gene is known to drive gene expression in both rods and cones (Liou et al., 1991; Yokoyama et al., 1992). The second, the mouse opsin promoter (MOP), has been shown to drive very high levels of gene expression in rods with some minimal basal activity in cones, and was chosen because the primary early defect in rds+/− mice occurs in rods (Flannery et al., 1997; Quiambao et al., 1997).

  • CHX10 targets a subset of photoreceptor genes

    2006, Journal of Biological Chemistry
    Citation Excerpt :

    DNA recovered from ChIP samples was subjected to real time PCR. The primers were designed for regions upstream of Rod arrestin, Rhodopsin, Red/green cone opsin, and Interphotoreceptor retinoid-binding protein (Irbp) (Fig. 3B), all of which are known to contain homeodomain binding sites (35–38). For negative controls, we amplified the β-globin promoter and the 3′ end of all the retinal target genes we analyzed.

  • Endogenous CRX expression and IRBP promoter activity in retinoblastoma cells

    2001, Brain Research
    Citation Excerpt :

    IRBP protein is detected only in photosensitive tissues [23] and its mRNA is detected uniquely in photoreceptor cells and pinealocytes [20,21]. The 5′ flanking sequence of the IRBP gene directs promoter activity only in the retina and pineal [3,16]. A short fragment, −123 to +18 relative to transcription start, of the IRBP gene is sufficient to direct retina-specific expression in transgenic mice [5,10].

  • A PDE6A promoter fragment directs transcription predominantly in the photoreceptor

    2001, Biochemical and Biophysical Research Communications
View all citing articles on Scopus
*

This study was supported in part by grants from the National Eye Institute (EY03829 to G. I. L. and EY06762 to P. A. O.), Howard Hughes Medical Institute (P. A. O.), and by an Unrestricted Departmental Award from Research to Prevent Blindness, Inc. to the Department of Ophthalmology, Medical College of Georgia.

View full text