(精选)microRNA Expression Levels, in Both Tissue and Blood, Correlate with Disease State in a Mouse Model of Pancreatic Cancer教学课件.pptVIP

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2018-03-21 发布于天津
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(精选)microRNA Expression Levels, in Both Tissue and Blood, Correlate with Disease State in a Mouse Model of Pancreatic Cancer教学课件.ppt

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Life Science Symposium Katie Freeman Conditionally expressed allele Genetic elements that inhibit transcription and translation flanked by LoxP sites Lox-STOP-Lox inserted upstream of modified genomic KRAS with Glycine 12 to Aspartic acid activating mutation. (Results in constitutive downstream signaling of ras pathway) Expression of mutated allele is achieved by interbreeding with animals that express Cre recombinase from pancreatic specific promoter PDX-1 or p48 P48 is required to commit cells to pancreatic fate. Restricted to acinar cells pDX expession is confined to islet cells. Maintains endogenous expression levels of KRS and Trp53 Excision of silencing cassette (LSL) occurs in pancreatic progenitor cells early in development and in the islet cells or acinar cells of adult mouse Results in expression throughout pancreas. Pancreas is only organ showing excision in DNA Are these microRNAs present in plasma? And if so do the levels correlate with the levels seen in tissue. Looked at the panel of 180 microRNAs that were available from Applied Biosystems at the time. 24 significantly upregulated vs control See increased expression correlating with increased in severity of disease state. A 2X then 6X up with progressive disease E PanIN 5X down F 3 fold down in PanIN, 6 fold down in PDA * microRNA Expression Levels, in Both Tissue and Blood, Correlate with Disease State in a Mouse Model of Pancreatic Cancer Katie B. Freeman PhD LifeScience Regional Technology Symposium September 29, 2009 microRNA Small, 19-23 nucleotide, naturally occurring regulatory RNAs Transcribed by pol II Non-protein coding Processed from longer hairpin RNAs First discovered in 1993 in C.elegans by V. Ambros and G. Ruvkun First mammalian microRNAs discovered in 2001 microRNAs regulate gene expression by binding to 3’UTR of mRNA and inhibiting translation miRNA Biogenesis and Maturation mRNA translation inhibition mRNA cleava