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Characterization of Celf1 function in mouse lens cell lines (746.1)
Author(s) -
Dang Christine,
Siddam Archana,
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.746.1
Subject(s) - gene knockdown , biology , gene , rna , lens (geology) , microbiology and biotechnology , small hairpin rna , rna binding protein , genetics , paleontology
Although the human genome encodes ~1500 RNA binding protein (RBP) encoding genes, mutations in only ~20 are associated with human disease. We used a novel bioinformatics approach iSyTE (integrated Systems Tool for Eye gene discovery) to identify a new RBP, Celf1 ( C UGBP E lav (embryonic lethal, abnormal vision)‐ L ike F amily member 1), in the lens that is potentially associated with cataract. Celf1 contains three conserved RNA Regulatory Motifs that function in sequence‐specific binding to target RNAs. To develop a new resource to study Celf1 function in lens cells, we generated and characterized Celf1 ‐knockdown (KD) mouse lens epithelial cell lines. We first selected the mouse lens epithelial cell line 21EM15 as it expresses several critical lens‐expressed genes and thus recapitulates important aspects of lens biology. We infected 21EM15 cells with lentiviral particles carrying inhibitory short hairpin (sh)RNAs against mouse Celf1 mRNA and isolated infected cells based on their acquired resistance to puromycin. Celf1 ‐KD was confirmed by reduced Celf1 expression in Western blotting. We are presently characterizing the effect of Celf1‐KD on distinct classes of RNA granules ( e.g. Stress granules and Processing bodies) involved in post‐transcriptional regulation. In sum, we describe development of a new Celf1‐KD lens cell line as a unique resource for gaining novel functional insights of an RBP in lens biology. Grant Funding Source : Supported by Pew Charitable Trusts