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Expression of mouse small interfering RNAs in lettuce using artificial microRNA technology.
Abstract Artificial miRNA technology enables the generation of siRNAs to regulate the expression of targeted genes. However, the application of siRNAs to alter gene expression is challenging due to their instability and requires a means to efficiently deliver siRNAs into the host. Here, we report that the siRNAs targeted to animal mRNAs can be heterologously expressed and stably produced in lettuce. We have modified rice miRNA precursors to produce siRNAs in lettuce with the potential to target mRNAs of mouse complement 3 (C3) and coagulation factor 7 (CF7). Expression of primary and mature siRNAs in the transge...
Source: BioTechniques - January 14, 2020 Category: Biotechnology Authors: Kakeshpour T, Tamang TM, Park WD, Manohar M, Yang J, Hirschi KD, Park S Tags: Biotechniques Source Type: research

Alfalfa dwarf cytorhabdovirus P protein is a local and systemic RNA silencing supressor which inhibits programmed RISC activity and prevents transitive amplification of RNA silencing.
In this study, we show that the phosphoprotein (P) encoded by the negative-sense RNA virus alfalfa dwarf virus (ADV), a species of the genus Cytorhabdovirus, family Rhabdoviridae, is a suppressor of RNA silencing. ADV P has a relatively weak local RSS activity, and does not prevent siRNA accumulation. On the other hand, ADV P strongly suppresses systemic RNA silencing, but does not interfere with the short-distance spread of silencing, which is consistent with its lack of inhibition of siRNA accumulation. The mechanism of suppression appears to involve ADV P binding to RNA-induced silencing complex proteins AGO1 and AGO4 a...
Source: Virus Research - August 15, 2016 Category: Virology Authors: Bejerman N, Mann KS, Dietzgen RG Tags: Virus Res Source Type: research

Cytorhabdovirus phosphoprotein shows RNA silencing suppressor activity in plants, but not in insect cells.
Abstract RNA silencing in plants and insects provides an antiviral defense and as a countermeasure most viruses encode RNA silencing suppressors (RSS). For the family Rhabdoviridae, no detailed functional RSS studies have been reported in plant hosts and insect vectors. In agroinfiltrated Nicotiana benthamiana leaves we show for the first time for a cytorhabdovirus, lettuce necrotic yellows virus (LNYV), that one of the nucleocapsid core proteins, phosphoprotein (P) has relatively weak local RSS activity and delays systemic silencing of a GFP reporter. Analysis of GFP small RNAs indicated that the P protein did no...
Source: Virology - January 12, 2015 Category: Virology Authors: Mann KS, Johnson KN, Dietzgen RG Tags: Virology Source Type: research