scholarly journals Gene expression evidence for off-target effects caused by RNA interference-mediated gene silencing of Ubiquitin-63E in the cattle tick Rhipicephalus microplus

2011 ◽  
Vol 41 (9) ◽  
pp. 1001-1014 ◽  
Author(s):  
A.E. Lew-Tabor ◽  
S. Kurscheid ◽  
R. Barrero ◽  
C. Gondro ◽  
P.M. Moolhuijzen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Rna Interference ◽  
Gene Silencing ◽  
Rhipicephalus Microplus ◽  
Cattle Tick ◽  
Expression Evidence ◽  
Target Effects

scholarly journals Recent Advances in the Development of Exogenous dsRNA for the Induction of RNA Interference in Cancer Therapy

Molecules ◽  
2021 ◽  
Vol 26 (3) ◽  
pp. 701
Author(s):  
Tatiana S. Golubeva ◽  
Viktoria A. Cherenko ◽  
Konstantin E. Orishchenko
Keyword(s):  
Gene Expression ◽  
Rna Interference ◽  
Molecular Biology ◽  
Gene Silencing ◽  
Regulation Of Gene Expression ◽  
Research Area ◽  
Disease Treatment ◽  
Practical Applications ◽  
Agricultural Plants ◽  
Insect Reproduction

Selective regulation of gene expression by means of RNA interference has revolutionized molecular biology. This approach is not only used in fundamental studies on the roles of particular genes in the functioning of various organisms, but also possesses practical applications. A variety of methods are being developed based on gene silencing using dsRNA—for protecting agricultural plants from various pathogens, controlling insect reproduction, and therapeutic techniques related to the oncological disease treatment. One of the main problems in this research area is the successful delivery of exogenous dsRNA into cells, as this can be greatly affected by the localization or origin of tumor. This overview is dedicated to describing the latest advances in the development of various transport agents for the delivery of dsRNA fragments for gene silencing, with an emphasis on cancer treatment.

scholarly journals Assessing the Functionality of RNA Interference (RNAi) in the Phloem-feeding Maize pest Dalbulus maidis

2021 ◽  
Author(s):  
Tara-Kay L. Jones ◽  
Julio S. Bernal ◽  
Raul F. Medina
Keyword(s):  
Gene Expression ◽  
Rna Interference ◽  
Gene Silencing ◽  
Pest Control ◽  
Messenger Rna ◽  
Specific Gene ◽  
Insect Vector ◽  
Dalbulus Maidis ◽  
Spiroplasma Kunkelii ◽  
Phloem Feeding

Dalbulus maidis [(DeLong & Wolcott), corn leafhopper], a phloem-feeding insect, is the most efficient vector of maize stunting pathogens (Spiroplasma kunkelii, Maize bushy stunt phytoplasma, and Maize rayado fino virus) in the Americas. Studies involving gene editing in insects are rapidly providing information that can potentially be used for insect vector and plant disease control. RNA interference (RNAi), a sequence-specific gene silencing method, is one of the most widely used molecular tools in functional genomics studies. RNAi uses exogenous double-stranded RNA (dsRNA) or small interfering RNA (siRNA) to prevent the production of proteins by inhibiting the expression of their corresponding messenger RNA (mRNA). In this study, we measured the efficacy of gene silencing, and its effects on D. maidis mortality as proof of concept that RNAi is a viable tool for use in genetic pest control of phloem-feeding insects. Oral delivery of dsRNA using an artificial diet was used to silence two key insect genes, vacuolar ATP synthase subunit B, and subunit D (V-ATPase B and V-ATPase D). Our results showed reduced gene expression of V-ATPase B and V-ATPase D after ingestion of dsRNA, and significantly higher mortality, and wing deformation, associated with reduced gene expression, compared to control insects that were not orally fed dsRNA. These results reveal RNAi as a viable tool for use in genetic pest control of phloem-feeding insects, and a way for further functional genomic studies, such as identification of potential target genes for either population suppression or population replacement of this vector of maize diseases.

scholarly journals Cross-Kingdom Small RNAs Among Animals, Plants and Microbes

Cells ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 371 ◽  
Author(s):  
Zeng ◽  
Gupta ◽  
Jiang ◽  
Yang ◽  
Gong ◽  
...  
Keyword(s):  
Gene Expression ◽  
Rna Interference ◽  
Gene Silencing ◽  
Horizontal Transfer ◽  
Small Rnas ◽  
Signal Transmission ◽  
Fungal Diseases ◽  
Small Interfering Rnas ◽  
Eukaryotic Species ◽  
Non Coding Rnas

Small RNAs (sRNAs), a class of regulatory non-coding RNAs around 20~30-nt long, including small interfering RNAs (siRNAs) and microRNAs (miRNAs), are critical regulators of gene expression. Recently, accumulating evidence indicates that sRNAs can be transferred not only within cells and tissues of individual organisms, but also across different eukaryotic species, serving as a bond connecting the animal, plant, and microbial worlds. In this review, we summarize the results from recent studies on cross-kingdom sRNA communication. We not only review the horizontal transfer of sRNAs among animals, plants and microbes, but also discuss the mechanism of RNA interference (RNAi) signal transmission via cross-kingdom sRNAs. We also compare the advantages of host-induced gene silencing (HIGS) and spray-induced gene silencing (SIGS) technology and look forward to their applicable prospects in controlling fungal diseases.

scholarly journals Global Effects on Gene Expression in Fission Yeast by Silencing and RNA Interference Machineries

2005 ◽  
Vol 25 (2) ◽  
pp. 590-601 ◽  
Author(s):  
Klavs R. Hansen ◽  
Gavin Burns ◽  
Juan Mata ◽  
Thomas A. Volpe ◽  
Robert A. Martienssen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Rna Interference ◽  
Gene Silencing ◽  
Fission Yeast ◽  
Histone Deacetylases ◽  
Zinc Finger Protein ◽  
Histone Deacetylation ◽  
Repeated Sequences ◽  
General Mechanism ◽  
Long Terminal Repeats

ABSTRACT Histone modifications influence gene expression in complex ways. The RNA interference (RNAi) machinery can repress transcription by recruiting histone-modifying enzymes to chromatin, although it is not clear whether this is a general mechanism for gene silencing or whether it requires repeated sequences such as long terminal repeats (LTRs). We analyzed the global effects of the Clr3 and Clr6 histone deacetylases, the Clr4 methyltransferase, the zinc finger protein Clr1, and the RNAi proteins Dicer, RdRP, and Argonaute on the transcriptome of Schizosaccharomyces pombe (fission yeast). The clr mutants derepressed similar subsets of genes, many of which also became transcriptionally activated in cells that were exposed to environmental stresses such as nitrogen starvation. Many genes that were repressed by the Clr proteins clustered in extended regions close to the telomeres. Surprisingly few genes were repressed by both the silencing and RNAi machineries, with transcripts from centromeric repeats and Tf2 retrotransposons being notable exceptions. We found no correlation between repression by RNAi and proximity to LTRs, and the wtf family of repeated sequences seems to be repressed by histone deacetylation independent of RNAi. Our data indicate that the RNAi and Clr proteins show only a limited functional overlap and that the Clr proteins play more global roles in gene silencing.

scholarly journals CRISPR therapy towards an HIV cure

2019 ◽  
Vol 19 (3) ◽  
pp. 201-208 ◽  
Author(s):  
Elena Herrera-Carrillo ◽  
Zongliang Gao ◽  
Ben Berkhout
Keyword(s):  
Gene Expression ◽  
Rna Interference ◽  
Optimal Design ◽  
Therapeutic Strategy ◽  
Genetic Diseases ◽  
High Specificity ◽  
Hiv Cure ◽  
Delivery Methods ◽  
Anti Hiv ◽  
Target Effects

Abstract Tools based on RNA interference (RNAi) and the recently developed clustered regularly short palindromic repeats (CRISPR) system enable the selective modification of gene expression, which also makes them attractive therapeutic reagents for combating HIV infection and other infectious diseases. Several parallels can be drawn between the RNAi and CRISPR-Cas9 platforms. An ideal RNAi or CRISPR-Cas9 therapeutic strategy for treating infectious or genetic diseases should exhibit potency, high specificity and safety. However, therapeutic applications of RNAi and CRISPR-Cas9 have been challenged by several major limitations, some of which can be overcome by optimal design of the therapy or the design of improved reagents. In this review, we will discuss some advantages and limitations of anti-HIV strategies based on RNAi and CRISPR-Cas9 with a focus on the efficiency, specificity, off-target effects and delivery methods.

scholarly journals RNA Interference and mRNA Silencing, 2004: How Far Will They Reach?

2004 ◽  
Vol 15 (2) ◽  
pp. 407-410 ◽  
Author(s):  
Thoru Pederson
Keyword(s):  
Gene Expression ◽  
Rna Interference ◽  
Gene Silencing ◽  
Rna Silencing ◽  
Regulation Of Gene Expression ◽  
Clinical Impact ◽  
Posttranscriptional Gene Silencing ◽  
Highly Effective ◽  
The Future ◽  
Recent Developments

The discoveries of RNA interference and RNA-mediated posttranscriptional gene silencing have opened an unanticipated new window on the regulation of gene expression as well as a facile and highly effective tool for knocking down gene expression in many organisms and cells. In addition, RNA interference and RNA silencing may conceivably be exploited for human therapeutics sometime in the future, possibly bringing greater clinical impact than have the so far disappointing antisense endeavors. This essay summarizes recent developments and offers some personalized perspectives, with emphasis on what we do not yet know.

Gene silencing to induce pathogen-derived resistance in plants.

2021 ◽  
pp. 4-13
Author(s):  
Elena Zuriaga ◽  
Ángela Polo-Oltra ◽  
Maria Luisa Badenes
Keyword(s):  
Rna Interference ◽  
Gene Silencing ◽  
Basic Research ◽  
Rnai Technology ◽  
History Of ◽  
Pathogen Derived Resistance ◽  
Target Effects

Abstract This chapter provides information on the history of the use of pathogen-derived resistance (PDR) in plants and use of PDR for basic research and commercial purposes. Some limitations of the RNA interference (RNAi) technology are presented and various tools that can be used to design RNAi constructs and screen for potential off-target effects are also discussed.

scholarly journals Pyrethroid Acaricide Resistance Is Proportional to P-450 Cytochrome Oxidase Expression in the Cattle Tick Rhipicephalus microplus

2018 ◽  
Vol 2018 ◽  
pp. 1-6 ◽  
Author(s):  
Raquel Cossío-Bayúgar ◽  
Francisco Martínez-Ibañez ◽  
Hugo Aguilar-Díaz ◽  
Estefan Miranda-Miranda
Keyword(s):  
Gene Expression ◽  
Pyrethroid Resistance ◽  
Linear Regression Analysis ◽  
Rhipicephalus Microplus ◽  
Cattle Tick ◽  
Synthetic Pyrethroids ◽  
Acaricide Resistance ◽  
Expression Levels ◽  
Field Isolates ◽  
Moderate Resistance

The goal of the present study was to assess the gene expression of xenobiotic metabolizing enzymes (XMEs) Cytochrome P-450 (CYP) and carboxylesterase (CE) related to detoxification of synthetic pyrethroids, plus acetylcholinesterase (AChE), in field isolates of acaricide-resistant Rhipicephalus microplus. The XMEs expression levels were assessed by mRNA measurement using quantitative reverse transcription PCR. The XME expression levels of field-isolated acaricide-resistant ticks were compared against acaricide-susceptible reference ticks used in this study as a gene expression baseline and represented as relative expression units (REU). Field isolates were subjected to toxicological bioassays and determined resistant to all the Pyr acaricides (Pyr), whereas most of them were found susceptible to organophosphorous acaricides (OP), with the exception of three isolates, which exhibited moderate resistance to Diazinon. Significantly higher levels of CYP were detected in pyrethroid-resistance ticks when compared to Su ticks (P<0.01). A linear regression analysis showed that pyrethroid acaricide resistance levels of R. microplus were proportional to the CYP expression levels (correlation coefficient (R):0.85; P<0.05). Analysis on CE expression levels showed only one isolate resistant to Pyr and OP with a statistically significant increase (P<0.01). AChE expression levels showed statistically significant (P<0.01) subexpression in all tick isolates when compared to the susceptible reference. Our results suggest that pyrethroid acaricide resistance in the cattle tick may be diagnosed by measuring the CYP expression levels using quantitative PCR.

scholarly journals Development of a cricket paralysis virus-based system for inducing RNA interference-mediated gene silencing in Diaphorina citri

2020 ◽  
Author(s):  
Emilyn E. Matsumura ◽  
Jared C. Nigg ◽  
Elizabeth M. Henry ◽  
Bryce W. Falk
Keyword(s):  
Gene Expression ◽  
Rna Interference ◽  
Gene Silencing ◽  
Plant Pathogens ◽  
Rnai Pathway ◽  
Insect Vector ◽  
Diaphorina Citri ◽  
Insect Virus ◽  
Viral Mutant ◽  
Paralysis Virus

AbstractDiaphorina citri, the Asian citrus psyllid, is the insect vector of the phloem-limited bacterium ‘Candidatus Liberibacter asiaticus’, which causes the most devastating citrus disease worldwide: Huanglongbing (HLB). An efficient cure for HLB is still not available and the management of the disease is restricted to the use of pesticides, antibiotics and eradication of infected plants. Plant- and insect-infecting viruses have attracted increasing attention for their potential to manipulate traits in insects, especially insect vectors of plant pathogens. However, so far there are no insect virus-based vectors available for use in D. citri. Cricket paralysis virus (CrPV) is a well-studied insect-infecting dicistrovirus with a wide host range and has been used as a model in previous translational studies. In this work, we demonstrate for the first time that CrPV is infectious and pathogenic to D. citri. We show that specific amino acid mutations in the CrPV primary cleavage DvExNPGP motif resulted in a viral mutant that was attenuated compared to wild-type CrPV during infection of either Drosophila cells line or adult D. citri insects. This attenuated CrPV mutant was then used as the backbone for engineering a recombinant CrPV-based vector to specifically alter D. citri gene expression via the RNA interference (RNAi) pathway, a technology called Virus Induced Gene Silencing (VIGS). As proof-of-concept, we engineered recombinant CrPV-based vectors carrying nucleotide sequences derived from a previously reported D. citri target gene: the inhibitor of apoptosis gene (IA). RT-qPCR analysis of insects either microinjected or fed with the recombinant CrPV mutants showed decreased IA gene expression as soon as viral replication was detected, indicating that the engineered CrPV-based VIGS system enables functional gene silencing in D. citri. This novel insect virus-based tool is easily amenable to genomic modification and represents a technical advance for understanding interactions between insect virus-based VIGS systems and D. citri.

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