(精选)MicroRNA教学课件.pptVIP

NEJM 353;17 0CT 2005 microchip analysis and Northern blotting showed that CLL cells from both patients had a substantial reduction in the expression of miR-16-1 as compared with that of normal CD5+ cells In Vitro Confirmation that mutations in genomic DNA encoding miRNA affects miRNA expression Cloned genomic sequence encoding both miR-16-1 and miR-15a (both wt and mutant C-T +7) Ligated into expression vector pSR-GFP-Neo Transfected (lipofectamine) into 293 cells Assessed miRNA expression by northern NEJM 353;17 0CT 2005 NEJM 353;17 0CT 2005 To identify a possible molecular effect of the C-T substitution, vectors containing either the wild-type allele of the miR-15a–miR-16-1 cluster or the mutated allele were prepared. The 293 cells, were transfected with the vectors. Significant association between the expression of certain microRNAs and the expression of ZAP-70, the mutational status of IgVH, and the time between diagnosis and initial treatment. A unique 13-gene molecular signature is associated with each prognostic factor. microRNA expression can be included in the markers with prognostic significance in CLL. microRNA signature may be relevant to the pathogenesis of CLL. Mechanism active in CLL pathogenesis?miR-15a and miR-16-1 induce apoptosis by targeting bcl2 Bcl2 protein expression is inversely correlated with miR-15a and miR-16-1 expression in CLL samples Bcl2 mRNA is a direct target of miR-15a and miR-16-1 miR-15a or miR-16-1 induce apoptosis in bcl2+ CLLcell line, while mutant miR-16-1 c-T+7 does not Cimmino PNAS 102:13944, 2005 Gene therapy and RNA interference Gene therapy, intends to provide therapeutic merit by introducing genetic material (DNA or RNA) encoding a protein that is missing or defective into a patient’s cells or tissues. A hallmark of gene therapy is the efficient delivery of these nucleic acids via the use of shuttle vectors Either nonviral, such as liposomes or nanoparticles, or derived from genetically modifi