Purchase this article with an account.
R Farjo, MI Othman, J Yu, SP MacNee, AJ Mears, S Yoshida, A Swaroop; Defining Mouse Eye Gene Expression Profiles with I-Gene Microarrays: A Resource for Vision Research . Invest. Ophthalmol. Vis. Sci. 2002;43(13):1427.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Purpose: To generate mouse microarrays of eye-expressed genes for developing gene expression profiles of eye development and disease. Methods: Initial efforts have focused on the production of mouse cDNA arrays that contain a comprehensive set of eye-expressed genes as well as a number of uncharacterized eye-expressed ESTs. Two cDNA libraries have been generated from mouse eye (E15.5 and P2.5) and a third from adult retina. In addition, libraries have been generated from Nrl-knockout retinae that have only cones and no rods. Over 7500 expressed sequence tags have been identified so far. Using the Chipwriter Pro robot (Virtek), more than 6500 cDNAs are spotted in duplicate onto glass slides. Microarray hybridizations are performed against a common reference RNA, which allows global comparison of all datasets generated with mouse I-Gene arrays. Results: A number of projects have been initiated in the lab or in collaboration with different investigators to define gene expression profiles of mutant mice, including models of retinal degeneration. The validity of the data is confirmed by replicate experiments. The expression profiles are being used to develop novel methods for normalization and image extraction. Significant changes in gene expression are verified with real-time RT-PCR. We will present optimal methods for array design, generation, hybridization and analysis. Conclusions: Defining the differential expression of genes in a given tissue or cell type can be readily accomplished with cDNA microarray technology. I-Gene arrays are proving to be an invaluable resource for generation of gene expression profiles of mutant mice. Our collection of mouse ESTs is expanding with the continued analysis of new mouse cDNA libraries. In conjunction with other gene expression tools, such as Affymetrix Genechips, SAGE and real-time RT-PCR, these arrays can give insights into the underlying variations that cause and occur with disease and assist in identifying potential targets for therapy. I-Gene arrays are available to interested vision scientists.
This PDF is available to Subscribers Only