(精选)Novel rules for microRNA教学课件.pptVIP

* * * Cellular Targets of EBV encoded microRNAs are enriched in genes involved in Apoptosis and Tumour Supression *Protein Interactions are from Human Protein Interaction map (HiMap) Scaria et al, Cell Microbiology 2007 Scaria and Jadhav, Retrovirology 2007 hp XC 3000 Cluster 288 Nodes Infiniband Interconnect 4.7 Teraflops Human miRNAs Genome Sequences | 3’UTR sequences Consensus Targets miRanda RNAhybrid TargetScan Large Scale Computation in 288 node 4 TeraFlop Supercomputer Computational pipeline for microRNA target prediction http://miracle.igib.res.in TargetmiR: Features microRNA Details and Validation Methods Interface Validated Targets Predicted Targets http://miracle.igib.res.in SCORING MATRIX Human 3’UTRdb Seed Counts db Seed Region 3’UTR Artifical miRNA (amiRNA) Artificially designed microRNAs (amiRNAs) HIV-1 transcripts targeted by amiRNAs miranda rna22 mirtif rnahybrid miracle amiRNA-001 GAG X amiRNA-002 GAG X amiRNA-003 ENV amiRNA-004 ENV amiRNA-005 POL amiRNA-006 POL amiRNA-007 POL X amiRNA-008 POL amiRNA-009 POL Computational Validation of Design HIV Genome Ultra-Conserved Regions Computational Design Scaria et al, Cell Microbiol. 2007;9(12):2784-2794 Design of artificial antiviral microRNAs (amiRNAs) in vitro validation of artificial miRNA CMV Luc SV 40 poly A signal. Target sequence. CMV Pre-miRNA SV 40 poly A signal. pMir reporter. pSilencer. The target sequence was cloned into the vector after the luciferase gene to form a fusion transcript (pmiR-reporter) and miRNA expression vector (pSilencer) where pre-miRNA were cloned. The luciferase activity would be decreased by binding of miRNAs to the 3 UTR of Firefly luciferase gene. Construct map of the plasmids used for the luciferase assay Luciferase activity of the reporter gene in the absence or presence of the amiR-01, amiR-04 and amiR-06 or either of reporter vector or miRNA expression vector shuffled measured. 293T cells were co-transfected with both the reporter gene and miRNA ex