Objective The aim of this review was to assess RNA interference (RNAi) and its possibility as a potential and powerful tool to develop highly specific double-stranded RNA ( dsRNA) or small interfering RNA (siRNA) base...Objective The aim of this review was to assess RNA interference (RNAi) and its possibility as a potential and powerful tool to develop highly specific double-stranded RNA ( dsRNA) or small interfering RNA (siRNA) based gene-silencing therapeutics. Data sources The data used in this review were obtained from the current RNAi-related research reports. Study selection dsRNA-mediated RNAi has recently emerged as a powerful reverse genetic tool to silence, gene expression in multiple organisms. The discovery that synthetic duplexes of 21 nucleotides siRNAs trigger gene-specific silencing in mammalian cells has further expanded the utility of RNAi in to the mammalian system. Data extraction The currently published papers reporting the discovery and mechanism of RNAi phenomena and application of RNAi on gene function in mammalian cells were included. Data synthesis Since the recent development of RNAi technology in the mammalian system, investigators have used RNAi to elucidate gene function, and to develop gene-based therapeutics by delivery exogenous siRNA or siRNA expressing vector. The general and sequence-specific inhibitory effects of RNAi that will be selective, long-term, and systemic to modulate gene targets mentioned in similar reports have caused much concern about its effectiveness in mammals and its eventual use as a therapeutic mordality. Conclusions It is certain that the ability of RNAi in mammals to silence specific genes, either when transfected directly as siRNAs or when generated from DNA vectors, will undoubtedly accelerate the study of gene function and might also be used as a potentially useful method to develop highly gene-specific therapeutic methods. It is also expected that RNAi might one day be used to treat human diseases.展开更多
Functional analysis for gene silencing suppres- sor of P14 gene of Beet necrotic yellow vein virus and S6 gene of Rice black streak dwarf virus was carried out by agro- in- filtration with recombinant vectors of Potat...Functional analysis for gene silencing suppres- sor of P14 gene of Beet necrotic yellow vein virus and S6 gene of Rice black streak dwarf virus was carried out by agro- in- filtration with recombinant vectors of Potato virus X. The phenotype observation of green fluorescent protein (GFP) expression and Northern blot showed that the gene silencing of gfp transgenic Nicotiana benthamiana induced by ho- mologous sequence was strongly suppressed by the immix- ture infiltration of either the P14 or the S6. In the suppressed plants, the gfp mRNA accumulation was higher than that in the non-suppressed controls and the symptoms caused by PVX infection became more severe, especially the gfp DNA methylation of plant genome was significantly inhabited when co-infiltrated with RBSDV S6 gene. These results sug- gested that these two virus genes were potentially to encode for proteins as RNA silencing suppressors.展开更多
植物通过控制细胞分裂和伸长决定器官的形状。为了研究器官形状决定的分子机理,通过EMS诱变分离得到一个叶形细长的拟南芥突变体。细胞生物学观察发现,该基因突变不仅影响了生长点中的细胞分裂,也影响了叶片细胞的形状和数目,其表皮细...植物通过控制细胞分裂和伸长决定器官的形状。为了研究器官形状决定的分子机理,通过EMS诱变分离得到一个叶形细长的拟南芥突变体。细胞生物学观察发现,该基因突变不仅影响了生长点中的细胞分裂,也影响了叶片细胞的形状和数目,其表皮细胞凸起数明显减少,呈单轴向伸长,因此将该突变体定名为slender leaves and cells(slc)。有趣的是,不同组织内细胞分裂和伸长受到不同程度的影响,说明SLC基因在协调细胞分裂和伸长过程中起关键作用。图位克隆结果表明,SLC与小RNA介导的基因沉默相关基因AGO1等位,其第574位组氨酸突变为酪氨酸。slc和ago1杂交F1代植物呈现突变体表型,证明AGO1和SLC确实为同一基因。以上结果表明,SLC/AGO1所介导的转录后基因沉默对控制植物器官和细胞形状决定均起重要作用。展开更多
文摘Objective The aim of this review was to assess RNA interference (RNAi) and its possibility as a potential and powerful tool to develop highly specific double-stranded RNA ( dsRNA) or small interfering RNA (siRNA) based gene-silencing therapeutics. Data sources The data used in this review were obtained from the current RNAi-related research reports. Study selection dsRNA-mediated RNAi has recently emerged as a powerful reverse genetic tool to silence, gene expression in multiple organisms. The discovery that synthetic duplexes of 21 nucleotides siRNAs trigger gene-specific silencing in mammalian cells has further expanded the utility of RNAi in to the mammalian system. Data extraction The currently published papers reporting the discovery and mechanism of RNAi phenomena and application of RNAi on gene function in mammalian cells were included. Data synthesis Since the recent development of RNAi technology in the mammalian system, investigators have used RNAi to elucidate gene function, and to develop gene-based therapeutics by delivery exogenous siRNA or siRNA expressing vector. The general and sequence-specific inhibitory effects of RNAi that will be selective, long-term, and systemic to modulate gene targets mentioned in similar reports have caused much concern about its effectiveness in mammals and its eventual use as a therapeutic mordality. Conclusions It is certain that the ability of RNAi in mammals to silence specific genes, either when transfected directly as siRNAs or when generated from DNA vectors, will undoubtedly accelerate the study of gene function and might also be used as a potentially useful method to develop highly gene-specific therapeutic methods. It is also expected that RNAi might one day be used to treat human diseases.
基金This work w as supported by the National Natural Science Founda-tion of China (Grants Nos.30325001 and 30471136).
文摘Functional analysis for gene silencing suppres- sor of P14 gene of Beet necrotic yellow vein virus and S6 gene of Rice black streak dwarf virus was carried out by agro- in- filtration with recombinant vectors of Potato virus X. The phenotype observation of green fluorescent protein (GFP) expression and Northern blot showed that the gene silencing of gfp transgenic Nicotiana benthamiana induced by ho- mologous sequence was strongly suppressed by the immix- ture infiltration of either the P14 or the S6. In the suppressed plants, the gfp mRNA accumulation was higher than that in the non-suppressed controls and the symptoms caused by PVX infection became more severe, especially the gfp DNA methylation of plant genome was significantly inhabited when co-infiltrated with RBSDV S6 gene. These results sug- gested that these two virus genes were potentially to encode for proteins as RNA silencing suppressors.
文摘植物通过控制细胞分裂和伸长决定器官的形状。为了研究器官形状决定的分子机理,通过EMS诱变分离得到一个叶形细长的拟南芥突变体。细胞生物学观察发现,该基因突变不仅影响了生长点中的细胞分裂,也影响了叶片细胞的形状和数目,其表皮细胞凸起数明显减少,呈单轴向伸长,因此将该突变体定名为slender leaves and cells(slc)。有趣的是,不同组织内细胞分裂和伸长受到不同程度的影响,说明SLC基因在协调细胞分裂和伸长过程中起关键作用。图位克隆结果表明,SLC与小RNA介导的基因沉默相关基因AGO1等位,其第574位组氨酸突变为酪氨酸。slc和ago1杂交F1代植物呈现突变体表型,证明AGO1和SLC确实为同一基因。以上结果表明,SLC/AGO1所介导的转录后基因沉默对控制植物器官和细胞形状决定均起重要作用。
