tailieunhanh - Báo cáo khoa hoc:" Shortcomings of short hairpin RNA-based transgenic RNA interference in mouse oocytes"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Shortcomings of short hairpin RNA-based transgenic RNA interference in mouse oocytes | Sarnova et al. Journal of Negative Results in BioMedicine 2010 9 8 http content 9 1 8 r M 1 JOURNAL OF NEGATIVE RESULTS IN BIOMEDICINE RESEARCH Open Access Shortcomings of short hairpin RNA-based transgenic RNA interference in mouse oocytes Lenka Sarnova1 2 Radek Malik1 Radislav Sedlacek2 Petr Svoboda1 Abstract Background RNA interference RNAi is a powerful approach to study a gene function. Transgenic RNAi is an adaptation of this approach where suppression of a specific gene is achieved by expression of an RNA hairpin from a transgene. In somatic cells where a long double-stranded RNA dsRNA longer than 30 base-pairs can induce a sequence-independent interferon response short hairpin RNA shRNA expression is used to induce RNAi. In contrast transgenic RNAi in the oocyte routinely employs a long RNA hairpin. Transgenic RNAi based on long hairpin RNA although robust and successful is restricted to a few cell types where long double-stranded RNA does not induce sequence-independent responses. Transgenic RNAi in mouse oocytes based on a shRNA offers several potential advantages including simple cloning of the transgenic vector and an ability to use the same targeting construct in any cell type. Results Here we report our experience with shRNA-based transgenic RNAi in mouse oocytes. Despite optimal starting conditions for this experiment we experienced several setbacks which outweigh potential benefits of the shRNA system. First obtaining an efficient shRNA is potentially a time-consuming and expensive task. Second we observed that our transgene which was based on a common commercial vector was readily silenced in transgenic animals. Conclusions We conclude that the long RNA hairpin-based RNAi is more reliable and cost-effective and we recommend it as a method-of-choice when a gene is studied selectively in the oocyte. Background RNA interference RNAi is a sequence-specific mRNA degradation induced by double stranded RNA dsRNA . Briefly long dsRNA is .