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Non-coding RNA Genes the underestimated players in an old game.

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Presentation on theme: "Non-coding RNA Genes the underestimated players in an old game."— Presentation transcript:

1 Non-coding RNA Genes the underestimated players in an old game

2 The estimated number of human genes keeps down down.  One goal of genome projects is to systematically identify genes.  100,000 to 21,000  the numerology based on three methods cDNA cloning and EST sequencing of polyadenylated mRNA identification of conserved coding exons by comparative genome analysis computational gene prediction

3 ncRNA genes has been underestimated  Non-coding RNA (ncRNA) genes  produce transcripts that function directly as structural, catalytic or regulatory RNA, rather than expressing mRNA that encode proteins.

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7 The regulation of DHRS4 in neuroblastoma cells is performed by a ncRNA produced by copy number variation of itself

8 NADPH-dependent retinol dehydrogenase/reductase Belongs to SDR family Firstly purified from the cytosole of rabbit liver by Huang,DY Highly catalytic capability of retinal reductase Expressed in many tissues of mammalian animal, maybe participates in a general metabolic pathway of retinoids. Molecular structure of NRDR

9 cloned from human liver, exists in various tissues. cloned from Hela cell. cervical squamous carcinoma tissues. selectively expressed in cervical squamous carcinoma tissues. cloned from neuroblastoma cells, exist in various carcinoma cell lines, may be functional ncRNA regulating DHRS4. DHRS4 may has copy number variation (CNV) between individuals.

10 Splicing forms in cervical squamous carcinoma: NRDRB1 Analyses of NRDRB1 mRNA splice variant. NRDR NRDRB1

11 A B C D E F Targeting of GFP-tagged NRDRB1 (A,B,C) and GFP-tagged NRDR (D,E,F) in HeLa cells.

12 SDS-PAGE and Western-blot analysis of NRDR and NRDRB1 expression after Immunoprecipitation.

13 Immunohistochemical staining of NRDRB1 and NRDR proteins in normal cervical epithelium and cervical squamous carcinoma tissues. A: Non-immune antibody control of normal cervical epithelium. B: Basal cells are positive for NRDR in normal cervical epithelium. C: Non-immune antibody control of cervical squamous carcinoma tissue. D: Only cells in the cancer nest region are positive for NRDRB1 in cervical squamous carcinoma tissue. A B C D

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