NDLI: Generation of Cre transgenic mice with postnatal RPE-specific ocular expression.

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Generation of Cre transgenic mice with postnatal RPE-specific ocular expression.

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Generation of Cre transgenic mice with postnatal RPE-specific ocular expression.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Iacovelli, Jared Wolkow, Natalie Zhao, Chen Ojha, Pallavi King, Ayala Gollomp, Kandace Dunaief, Joshua L. Esumi, Noriko Lukinova, Nina Zack, Donald J. Veldman, Peter Feiner, Leonard Vollrath, Douglas Pierce, Eric A.
Description	Country affiliation: United States Author Affiliation: Iacovelli J ( F. M. Kirby Center for Molecular Ophthalmology, Scheie Eye Institute, University of Pennsylvania, Philadelphia, Pennsylvania, USA.)
Abstract	PURPOSE: To generate and characterize a constitutively active, RPE-specific, cre-expressing transgenic mouse line. This line can be used to create RPE-specific knockouts by crossing with mice harboring loxP-flanked (floxed) genes. METHODS: A transgene construct was assembled with the BEST1 promoter driving cre expression. Transgenic mice were generated on a C57BL/6 background. Cre expression was assessed by immunofluorescence and Western blot analysis. Cre enzymatic activity was tested by crossing to three lines with floxed DNA regions and detecting deletion of the intervening sequences or through histochemical detection of lacZ activity. Potential cre-mediated toxicity was assessed by retinal histology up to 24 months of age and by electroretinography. RESULTS: The BEST1-cre line with expression in the highest percentage of RPE cells displayed a patchy mosaic expression pattern, with 50% to 90% of RPE cells expressing cre. In mice outcrossed to a mixed B6/129 background, expression was consistently found in 90% of RPE cells. Within the eye, only the RPE cells were immunoreactive with an anti-cre antibody. Maximum cre expression quantified by Western blot analysis occurred at P28. Crosses with three lines containing floxed sequences revealed RPE-specific cre activity in the eye and extraocular expression limited to the testes. Histology and electroretinography showed no cre-mediated RPE toxicity. CONCLUSIONS: This BEST1-cre transgenic line enables generation of RPE-specific knockout mice. The mosaic expression pattern provides an internal control; the non-cre-expressing RPE cells continue to express the floxed genes. These mice should facilitate study of the multifunctional RPE and the generation of mouse models of human retinal disease.
ISSN	01460404
e-ISSN	15525783
DOI	10.1167/iovs.10-6347
Journal	Investigative Opthalmology & Visual Science
Issue Number	3
Volume Number	52
Language	English
Publisher	Association for Research in Vision and Ophthalmology
Publisher Date	2011-03-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Gene Expression Regulation, Enzymologic Physiology Integrases Genetics Retinal Pigment Epithelium Enzymology Animals Blotting, Western Electroretinography Eye Proteins Fluorescent Antibody Technique, Indirect Ion Channels Mice Mice, Inbred C57bl Mice, Transgenic Polymerase Chain Reaction Promoter Regions, Genetic Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't Discipline Ophthalmology
Content Type	Text
Resource Type	Article
Subject	Ophthalmology Sensory Systems Cellular and Molecular Neuroscience

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