(精选)New Ago2-associated microRNAs identified by deep sequencing in human embryonic stem cells教学课件.pptVIP

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(精选)New Ago2-associated microRNAs identified by deep sequencing in human embryonic stem cells教学课件.ppt

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MicroRNAs and Stem Cells MicroRNAs and Stem Cells MicroRNAs and Stem Cells MicroRNAs and Stem Cells MicroRNAs and Stem Cells MicroRNAs and Stem Cells New Ago2-associated microRNAs identified by deep sequencing in human embryonic stem cells Ronald P. Hart Rutgers Stem Cell Research Center W.M. Keck Center for Collaborative Neuroscience Department of Cell Biology Neuroscience Rutgers University What are microRNAs and why are they important in stem cells? MicroRNAs are encoded by genome Up to 10,000 copies per genome (800 known) Selectively inhibit mRNA translation—multiple targets Small group of microRNAs are “ES-specific” Houbaviy, 2003 Production of mature microRNAs required for ES cell differentiation and “stemness” Kanellopoulou, 2005; Wang, 2007 Do regulated microRNAs regulate or modulate differentiation? Two examples Mir-302 family (includes miR-302a, miR-302b, miR-302c, miR-302d, miR-373, miR-520e, miR-520a-3p, miR-520b, miR-520c-3p, miR-520d-3p, miR-302e) Expression induced by Oct4/Sox4 Known targets include cyclin D1 Inhibition shifts cells towards G1 phase Addition of exogenous mir-302 enhances production of iPS cells Let-7 family (let-7a, let-7b, let-7c, let-7d, let-7f, miR-98, let-7g, let-7i) Constitutively transcribed, processing inhibited by lin28 (one of the proteins known to induce iPS) Lin28 rapidly drops upon differentiation, leading to processing of let7 Targets include Ras, c-myc, HMGA2, CDC25A and CDK6 Regulated microRNAs group hESC samples by phenotype and predict targeting by cross-correlation Predicted inhibition of g-1-laminin translation by stem-specific microRNAs Lakshmipathy et al., Stem Cells Devel. 2007 Search with “deep” sequencing Hypothesis: Regulated stem cell microRNAs help to direct gene regulation early in differentiation. Why limit detection to known small RNAs? Up to 10,000 microRNA-encoding loci predicted (Rigoutsos/IBM or Rosetta Genomics) Use deep sequencing, fit small RNA/genome alignments to computed model of Microproc