scholarly journals Involvement of Multiple Gene-Silencing Pathways in a Paramutation-like Phenomenon in Arabidopsis

Cell Reports ◽  
2015 ◽  
Vol 11 (8) ◽  
pp. 1160-1167 ◽  
Author(s):  
Zhimin Zheng ◽  
Hasi Yu ◽  
Daisuke Miki ◽  
Dan Jin ◽  
Qingzhu Zhang ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Multiple Gene ◽  
Silencing Pathways ◽  
Multiple Gene Silencing

Disulfide bond-conjugated dual PEGylated siRNAs for prolonged multiple gene silencing

Biomaterials ◽  
2013 ◽  
Vol 34 (28) ◽  
pp. 6930-6937 ◽  
Author(s):  
Zi-Xian Liao ◽  
Chun-Wen Hsiao ◽  
Yi-Cheng Ho ◽  
Hsin-Lung Chen ◽  
Hsing-Wen Sung
Keyword(s):  
Gene Silencing ◽  
Disulfide Bond ◽  
Multiple Gene ◽  
Multiple Gene Silencing

scholarly journals Multiple gene silencing in STAT pathway in K562 cells

2019 ◽  
Vol 4 ◽  
pp. 13-20
Author(s):  
Vinod Rajendran ◽  
Sudha S. Deo
Keyword(s):  
Gene Expression ◽  
Cell Proliferation ◽  
Gene Silencing ◽  
Cell Line ◽  
Quantitative Polymerase Chain Reaction ◽  
K562 Cells ◽  
Inhibition Of Proliferation ◽  
Multiple Gene ◽  
Proliferation And Apoptosis ◽  
Multiple Gene Silencing

Context: Chronic myeloid leukemia (CML) is characterized by the presence of a fusion oncoprotein BCR-ABL. This mutation imparts a constitutive phosphorylation activity of tyrosine residues in the cellular proteins. One of the targets of BCR-ABL is the STAT5 protein, which when phosphorylated induces gene expression of antiapoptotic proteins such as BCL-XL. The STAT pathway has been targeted in the past by disrupting any one protein only. A multiple gene silencing has never been done in this pathway. Aim: The aim of this study was to compare the effects of downregulation of BCR-ABL, STAT5A, STAT5B, and BCL-XL, individually and simultaneously, in human CML cell line (K562 cells) through RNA interference (RNAi). Further, gene expression, inhibition of proliferation, and apoptosis induction were assessed in K562 cells. Materials and Methods: K562 cells were transfected with various combinations of small iRNA (siRNA) and the expressions of aforesaid genes were determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot analysis. K562 cell proliferation and apoptosis were analyzed using 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide and flow cytometry, respectively. The results were compared through one-way analysis of variance. Results: qPCR and western blotting results post-siRNA transfection confirmed the targeted gene suppression and protein reduction in K562 cells. The cell proliferation assay and apoptosis assay revealed that simultaneous gene silencing of BCR-ABL, STAT5A, STAT5B, and BCL-XL had the highest killing effect on K562 cells as compared to knocking down these genes individually or in any other combinations. Conclusions: This was the first time it was shown that multiple gene silencing in STAT pathway in CML cell line K562 was better as compared to individual gene silencing.

DDX21 interacts with nuclear AGO2 and regulates the alternative splicing of SMN2

2020 ◽  
Vol 85 (2) ◽  
pp. 272-279
Author(s):  
Mengting Gong ◽  
Xi Zhang ◽  
Yaru Wang ◽  
Guiyan Mao ◽  
Yangqi Ou ◽  
...  
Keyword(s):  
Gene Expression ◽  
Alternative Splicing ◽  
Gene Silencing ◽  
Target Gene ◽  
Molecular Mechanisms ◽  
Regulation Of Gene Expression ◽  
Dependent Manner ◽  
Indirect Interaction ◽  
Multiple Gene ◽  
Multiple Gene Silencing

ABSTRACT AGO2 is the only member of mammalian Ago protein family that possesses the catalytic activity and plays a central role in gene silencing. Recently researches reported that multiple gene silencing factors, including AGO2, function in the nuclei. The molecular mechanisms of the gene silencing factors functioning in nuclei are conducive to comprehend the roles of gene silencing in pretranslational regulation of gene expression. Here, we report that AGO2 interacts with DDX21 indirectly in an RNA-dependent manner by Co-IP and GST-Pulldown assays and the 2 proteins present nuclei foci in the immunofluorescence experiments. We found that DDX21 up-regulated the protein level of AGO2 and participated in target gene, SNM2, alternative splicing involved in AGO2 by the indirect interaction with AGO2, which produced different transcripts of SMN2 in discrepant expression level. This study laid important experiment foundation for the further analysis of the nuclear functions of gene silencing components.

scholarly journals Simultaneous silencing of two different Arabidopsis genes with a novel virus-induced gene silencing vector

Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Kunxin Wu ◽  
Yadan Wu ◽  
Chunwei Zhang ◽  
Yan Fu ◽  
Zhixin Liu ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Double Mutant ◽  
Target Genes ◽  
Rna Binding ◽  
Protein Product ◽  
Antiviral Defense ◽  
Virus Induced Gene Silencing ◽  
Argonaute 2 ◽  
Plant Genes ◽  
Silencing Pathways

Abstract Background Virus-induced gene silencing (VIGS) is a useful tool for functional characterizations of plant genes. However, the penetrance of VIGS varies depending on the genes to be silenced, and has to be evaluated by examining the transcript levels of target genes. Results In this report, we report the development of a novel VIGS vector that permits a preliminary assessment of the silencing penetrance. This new vector is based on an attenuated variant of Turnip crinkle virus (TCV) known as CPB that can be readily used in Arabidopsis thaliana to interrogate genes of this model plant. A CPB derivative, designated CPB1B, was produced by inserting a 46 nucleotide section of the Arabidopsis PHYTOENE DESATURASE (PDS) gene into CPB, in antisense orientation. CPB1B induced robust PDS silencing, causing easily visible photobleaching in systemically infected Arabidopsis leaves. More importantly, CPB1B can accommodate additional inserts, derived from other Arabidopsis genes, causing the silencing of two or more genes simultaneously. With photobleaching as a visual marker, we adopted the CPB1B vector to validate the involvement of DICER-LIKE 4 (DCL4) in antiviral defense against TCV. We further revealed the involvement of ARGONAUTE 2 (AGO2) in PDS silencing and antiviral defense against TCV in dcl2drb4 double mutant plants. These results demonstrated that DOUBLE-STRANDED RNA-BINDING PROTEIN 4 (DRB4), whose protein product (DRB4) commonly partners with DCL4 in the antiviral silencing pathway, was dispensable for PDS silencing induced by CPB1B derivative in dcl2drb4 double mutant plants. Conclusions The CPB1B-based vector developed in this work is a valuable tool with visualizable indicator of the silencing penetrance for interrogating Arabidopsis genes, especially those involved in the RNA silencing pathways.

Phosphorylation of Argonaute proteins: regulating gene regulators

2008 ◽  
Vol 413 (3) ◽  
pp. e7-e9 ◽  
Author(s):  
Sabine Rüdel ◽  
Gunter Meister
Keyword(s):  
Gene Silencing ◽  
Translational Regulation ◽  
Mitogen Activated Protein Kinase ◽  
Rna Turnover ◽  
Processing Bodies ◽  
P Bodies ◽  
Biochemical Journal ◽  
Mitogen Activated Protein ◽  
Silencing Pathways ◽  
Protein Kinase Signalling

Members of the Ago (Argonaute) protein family are the mediators of small RNA-guided gene-silencing pathways including RNAi (RNA interference), translational regulation by miRNAs (microRNAs) and transcriptional silencing. Recent findings by Zeng et al. in this issue of the Biochemical Journal demonstrate that Ago proteins are post-translationally modified by phosphorylation of Ser387. Mutating Ser387 to alanine leads to reduced localization of human Ago2 to cytoplasmic P-bodies (processing bodies), cellular sites where RNA turnover and, at least in part, miRNA-guided gene regulation occurs. Zeng et al. further show that a member of the MAPK (mitogen-activated protein kinase) signalling pathway phosphorylates Ago2 at Ser387, suggesting that Ago2-mediated gene silencing might be linked to distinct signalling pathways.

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