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Identification of differentially expressed transcripts in Tdrd7 null mutant mouse lens (747.1)
Author(s) -
Patel Shaili,
Barnum Carrie,
Lachke Salil
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.747.1
Subject(s) - cataracts , lens fiber , biology , knockout mouse , mutant , embryonic stem cell , germline , lens (geology) , genetics , eye development , laser capture microdissection , microbiology and biotechnology , phenotype , gene knockout , gene , gene expression , paleontology
Lens is a transparent tissue within the eye that focuses light on the retina and facilitates high‐resolution vision. Cataract is an eye disease that results due to the loss of lens transparency and causes severely impaired vision. It can be caused by genetic changes, aging or physiological conditions like diabetes. Treatment depends upon the patient's specific visual needs and often involves cataract surgery. Over 77 million individuals are affected worldwide, and in the United States alone, costs exceed $3 billion annually. Recently, a mutations in the human TDRD7 (Tudor domain containing protein 7) gene were shown to cause congenital/pediatric cataracts in patients. Moreover, it was also demonstrated that Tdrd7 targeted germline knockout or ENU‐induced knockout mouse mutants exhibit cataracts that closely resemble the human phenotype. My research goal is to understand the regulatory function of Tdrd7 in the lens. Specifically, I aim to investigate if Tdrd7 deficiency affects the mRNA profiles in the apical and the basal regions of differentiating lens fiber cells. To achieve this I have expanded the Tdrd7 germline knockout mouse mutant colony and collected Tdrd7 mutant and control (Tdrd7+/‐ mice that do not exhibit cataracts) embryonic lens tissue for analysis. To study the expression of mRNAs in different locations within fiber cells, I have undertaken standardization of an approach using Laser Capture Microdissection (LCM). LCM on embryonic lens sections from Tdrd7 null mutants and control will be used to isolate the apical and the basal fiber cell regions at embryonic stage E10.5, E11.5, and E12.5. Once the specific tissues are collected, expression of mRNAs will be tested by whole genome expression profiling using Illumina WG‐6 microarrays. Comparative analysis of gene expression profiles of the apical and basal tissues from mutant and control will allow the identification of transcripts that are misregulated as a result of Tdrd7 mutation.