Mouse opsin promoter-directed Cre recombinase expression in transgenic mice.
OBJECTIVE
Gene inactivation with homologous recombination in mice is a widely used tool to study gene function. However, many proteins play essential roles in a number of tissues and germline gene inactivation often results in embryonic lethality. To overcome this limitation and to dissect the functions of essential genes beyond embryonic development, we generated mouse rod opsin promoter-controlled cre transgenic mice with a goal of obtaining transgenic lines with a range of Cre activity in rod photoreceptors.


METHODS
Transgenic mice expressing Cre recombinase directed by a long or short mouse opsin promoter were generated. Candidate Cre-expressing lines were identified with RT-PCR and Western blot analysis. Potentially useful Cre-expressing lines were characterized further with immunohistochemistry, PCR, and functional analysis using a Cre-activatable lacZ reporter mouse strain (R26R) to determine temporal and spatial patterns of Cre expression. Retinal function and morphology in these mouse lines were analyzed with electroretinography (ERG) and light microscopy of hematoxylin and eosin stained retinal sections.


RESULTS
Transgenic mice expressing Cre in rod photoreceptors were generated. Characterization of candidate photoreceptor-specific Cre mice using immunohistochemistry and functional assays demonstrated that an efficient Cre-mediated recombination occurred in rod photoreceptor cells in one mouse line and a mosaic Cre-mediated recombination occurred in rod photoreceptors and rod bipolar cells in another mouse line. Further analysis of these mice with ERG and morphological examination suggested that the retinas of eight-month-old adults were normal.


CONCLUSIONS
We have generated transgenic mice expressing Cre recombinase in rod photoreceptors. One transgenic mouse line was capable of carrying out efficient Cre-mediated recombination in rod photoreceptors. Another transgenic mouse line was capable of carrying out mosaic Cre-mediated recombination in rod photoreceptors and bipolar cells across the whole retina. These mice will be useful tools for Cre/lox-based gene activation and inactivation, as well as genetic mosaics, in rod photoreceptors and rod bipolar cells.
Le
Yun-Zheng
YZ
Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA. Yun-Le@ouhsc.edu
Zheng
Lixin
L
Zheng
Wei
W
Ash
John D
JD
Agbaga
Martin-Paul
MP
Zhu
Meili
M
Anderson
Robert E
RE
D000818:Animals; D015870:Gene Expression; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic / Q000235:genetics*; D009030:Mosaicism; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D012160:Retina / Q000166:cytology / Q000201:enzymology; D051245:Retinal Bipolar Cells / Q000201:enzymology; D017948:Retinal Rod Photoreceptor Cells / Q000201:enzymology*; D017299:Rod Opsins / Q000235:genetics*; D014018:Tissue Distribution
D017299:Rod Opsins; C045073:Cre recombinase; D019426:Integrases