RNA interference by short hairpin RNAs expressed in vertebrate cells
作者： Gregory J. Hannon , Douglas S. Conklin
DOI: 10.1385/1-59259-750-5:255
关键词: Genetics 、 RNA interference 、 RNA 、 Small interfering RNA 、 RNA silencing 、 Gene silencing 、 Biology 、 Cell biology 、 Gene expression 、 Small hairpin RNA 、 Trans-acting siRNA
摘要: RNA interference (RNAi) is now established as a general method to silence gene expression in variety of organisms. Double-stranded (dsRNA), when introduced cells, interferes with the homologous genes, disrupting their normal function. In mammals, transient delivery synthetic short interfering RNAs (siRNAs), which resemble processed form standard double stranded RNAi triggers, effective silencing mammalian genes. Issues related transfer efficiency and duration effect, however, restrict spectrum applications siRNAs mammals. These shortcomings have been solved by cellular hairpin (shRNAs) from DNA vectors. shRNAs are indistinguishable terms efficacy mechanism but can be produced within cells this way, shRNA makes possible creation continuous cell lines transgenic animals suppression target stably maintained RNAi. As result, types RNAi-based function analysis that carried out mammals greatly expanded. We describe methods for construction stable expressing vectors suitable generating loss alleles vitro or vivo.
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