High Density of Aligned Nanowire Treated with Polydopamine for Efficient Gene Silencing by siRNA According to Cell Membrane Perturbation

2016 ◽  
Vol 8 (29) ◽  
pp. 18693-18700 ◽  
Author(s):  
Baiju G. Nair ◽  
Kyoji Hagiwara ◽  
Motoki Ueda ◽  
Hsiao-hua Yu ◽  
Hsian-Rong Tseng ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Gene Silencing ◽  
High Density ◽  
Membrane Perturbation ◽  
Efficient Gene

scholarly journals A cassava common mosaic virus vector for virus-induced gene silencing in cassava

Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Decai Tuo ◽  
Peng Zhou ◽  
Pu Yan ◽  
Hongguang Cui ◽  
Yang Liu ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Mosaic Virus ◽  
Gene Function ◽  
Viral Vector ◽  
Function Analysis ◽  
Systemic Infection ◽  
Cdna Clones ◽  
Virus Induced Gene Silencing ◽  
Efficient Gene ◽  
Gene Function Analysis

Abstract Background Cassava is an important crop for food security and industry in the least-developed and developing countries. The completion of the cassava genome sequence and identification of large numbers of candidate genes by next-generation sequencing provide extensive resources for cassava molecular breeding and increase the need for rapid and efficient gene function analysis systems in cassava. Several plant virus-induced gene silencing (VIGS) systems have been developed as reverse genetic tools for rapid gene function analysis in cassava. However, these VIGS vectors could cause severe viral symptoms or inefficient gene silencing. Results In this study, we constructed agroinfection-compatible infectious cDNA clones of cassava common mosaic virus isolate CM (CsCMV-CM, genus Potexvirus, family Alphaflexiviridae) that causes systemic infection with mild symptoms in cassava. CsCMV-CM was then modified to a viral vector carrying the Nimble cloning frame, which facilitates the rapid and high-throughput cloning of silencing fragments into the viral genome. The CsCMV-based vector successfully silenced phytoene desaturase (PDS) and magnesium chelatase subunit I (ChlI) in different cassava varieties and Nicotiana benthamiana. The silencing of the ChlI gene could persist for more than two months. Conclusions This CsCMV-based VIGS system provides a new tool for rapid and efficient gene function studies in cassava.

High-throughput vectors for efficient gene silencing in plants

2002 ◽  
Vol 29 (10) ◽  
pp. 1217 ◽  
Author(s):  
Chris A. Helliwell ◽  
S. Varsha Wesley ◽  
Anna J. Wielopolska ◽  
Peter M. Waterhouse
Keyword(s):  
Gene Silencing ◽  
Insertional Mutagenesis ◽  
Single Step ◽  
Plant Genome ◽  
Specific Gene ◽  
Single Polymerase Chain Reaction ◽  
Efficient Gene ◽  
Gene Redundancy ◽  
Endogenous Genes ◽  
Rapid Generation

A major challenge in the post-genome era of plant biology is to determine the functions of all genes in the plant genome. A straightforward approach to this problem is to reduce or knockout expression of a gene with the hope of seeing a phenotype that is suggestive of its function. Insertional mutagenesis is a useful tool for this type of study but is limited by gene redundancy, lethal knockouts, non-tagged mutants, and the inability to target the inserted element to a specific gene. The efficacy of gene silencing in plants using inverted-repeat transgene constructs that encode a hairpin RNA (hpRNA) has been demonstrated by a number of groups, and has several advantages over insertional mutagenesis. In this paper we describe two improved pHellsgate vectors that facilitate rapid generation of hpRNA-encoding constructs. pHellsgate 4 allows the production of an hpRNA construct in a single step from a single polymerase chain reaction product, while pHellsgate 8 requires a two-step process via an intermediate vector. We show that these vectors are effective at silencing three endogenous genes in Arabidopsis, FLOWERING LOCUS C, PHYTOENE DESATURASE and ETHYLENE INSENSITIVE 2. We also show that a construct of sequences from two genes silences both genes.

scholarly journals Diblock Copolymer Hydrophobicity Facilitates Efficient Gene Silencing and Cytocompatible Nanoparticle-Mediated siRNA Delivery to Musculoskeletal Cell Types

2017 ◽  
Vol 18 (11) ◽  
pp. 3753-3765 ◽  
Author(s):  
Dominic W. Malcolm ◽  
Margaret A. T. Freeberg ◽  
Yuchen Wang ◽  
Kenneth R. Sims ◽  
Hani A. Awad ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Diblock Copolymer ◽  
Sirna Delivery ◽  
Cell Types ◽  
Efficient Gene

Lauroylated Histidine-Enriched S413-PV Peptide as an Efficient Gene Silencing Mediator in Cancer Cells

2020 ◽  
Vol 37 (10) ◽  
Author(s):  
Catarina M. Morais ◽  
Ana M. Cardoso ◽  
Luísa Aguiar ◽  
Nuno Vale ◽  
Clévio Nóbrega ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Cancer Cells ◽  
Efficient Gene

Efficient gene silencing induction in tomato by a viral satellite DNA vector

2007 ◽  
Vol 125 (2) ◽  
pp. 169-175 ◽  
Author(s):  
Xinzhong Cai ◽  
Changchun Wang ◽  
Youping Xu ◽  
Qiufang Xu ◽  
Zhong Zheng ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Satellite Dna ◽  
Efficient Gene ◽  
Dna Vector

scholarly journals Lipid utilization by human lymphocytes is correlated with high-density-lipoprotein binding site activity

1992 ◽  
Vol 285 (1) ◽  
pp. 105-112 ◽  
Author(s):  
Q Xu ◽  
G Jürgens ◽  
L A Huber ◽  
G Böck ◽  
H Wolf ◽  
...  
Keyword(s):  
Cell Membrane ◽  
High Density Lipoprotein ◽  
Binding Site ◽  
Human Peripheral Blood ◽  
Human Lymphocytes ◽  
Density Lipoprotein ◽  
Lipid Binding ◽  
High Density ◽  
Cell Protein ◽  
Significant Difference

The nature and physiological importance of high-density lipoprotein (HDL) binding sites on unstimulated (resting) and mitogen-activated (blast) human peripheral blood lymphocytes were investigated. Specific HDL binding on resting and blast T-lymphocytes was saturable at 50 micrograms of 125I-HDL/ml and of high affinity, with Kd values of 8.1 x 10(-8) M and 6.5 x 10(-8) M, respectively, and Bmax. values of 79 ng and 180 ng/mg of cell protein respectively at 4 degrees C. Binding of HDL double-labelled with fluorescent dioctadecylindocarbocyanine (Dil) and isotope (125I) as well as of single fluorescence- or isotope-labelled HDL was inhibited competitively by HDL apoproteins. Studies of the cholesterol flux between the cells and HDL showed that HDL, low-density lipoprotein (LDL) or BSA at a concentration of 100 micrograms/ml in the tissue culture medium did not result in a significant difference in exogenous [3H]cholesterol efflux from the cell membrane at 37 degrees C. Proliferating T-blasts incorporated more cholesterol from HDL or LDL than did resting lymphocytes. When the cells were pulsed with 125I-HDL and chased in fresh lipid-free medium, up to 80% of the radioactivity released was not precipitable with trichloroacetic acid. This percentage decreased in a competitive manner when unlabelled HDL was present in the chase incubation medium. Finally, cultivation of lymphocytes with conditioned medium from macrophages increased Dil-HDL binding/uptake, while it was decreased by mevinolin-induced inhibition of hydroxymethylglutaryl-coA reductase. In conclusion, human lymphocytes possess a HDL binding site (receptor) responsible for lipid binding/uptake and concomitant internalization and degradation of apoproteins from HDL, but not for reverse cell membrane cholesterol transport. The activity of the binding site is up-regulated during cell proliferation and down-regulated during cell growth suppression.

Comparison of approaches for efficient gene silencing induced by microRNA-based short hairpin RNA and indicator gene expression

2009 ◽  
Vol 37 (4) ◽  
pp. 1831-1839 ◽  
Author(s):  
Z. X. Shan ◽  
Q. X. Lin ◽  
C. Y. Deng ◽  
Z. L. Zhou ◽  
H. H. Tan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Silencing ◽  
Short Hairpin Rna ◽  
Hairpin Rna ◽  
Efficient Gene ◽  
Short Hairpin
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