(精选)Translational repression by miRNAs教学课件.pptVIP

stress induces CAT1-expression: 2.3. Transition to P-Bodies and a regulatory mechanism stress induces CAT1-expression independent of de-novo mRNA-synthesis: 2.3. Transition to P-Bodies and a regulatory mechanism stress induces CAT1-expression independent of de-novo mRNA-synthesis or miR122-degradation: 2.3. Transition to P-Bodies and a regulatory mechanism used constructs: 2.3. Transition to P-Bodies and a regulatory mechanism influence of 3′-UTR on reporter expression: 2.3. Transition to P-Bodies and a regulatory mechanism expression of reporter-constructs under stress: 2.3. Transition to P-Bodies and a regulatory mechanism Translational repression by miRNAs Talk by Stefan Kohl 1. Small Regulatory RNAs 2. Translational Repression 2.1. Initiation 2.2. Elongation 2.3. Transition to P-Bodies and a regulatory mechanism 3. Conclusion 4. Literatur 1. Small Regulatory RNAs 1. Small Regulatory RNAs involved in the regulation of developmental processes or tissue specific genexpression transcribed either from the same locus as the regulated mRNA (siRNA) or from distant loci in the genome (miRNA) can act on different (miRNA) or are specific for certain mRNAs (siRNA) 1. Small Regulatory RNAs pair perfectly (siRNA) or imperfectly in the 3′-UTR (miRNA) with their targets associated with different proteins (Ribonucleoprotein Particle - RNPs) estimates: ~ 4% of the humane genome code for miRNAs ~ 30% of human genes are regulated by miRNAs Chu Cy, Rana TM; PLoS Biology Vol. 4, No. 7 2. Translational Repression 2.1. Initiation 2.1. Repression of Translation Initiation Thermann and Hentze: transfection of D. melanogaster embryo extracts and cultured cells with reporter mRNAs (1 hr) qPCR and luciferase assays sucrose gradient analysis sedimentation after in-vitro transcription with cycloheximide: 2.1. Repression of Translation Initiation control + anti-miR2 + anti-let7 blue = FL-WT; red = FL-mut sedimentation aft