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Gene Silencing By Rna Interference Technology And Application Pdf

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Pocket K No. 34: RNAi for Crop Improvement

RNA interference RNAi , is a powerful new technology in the discovery of genetic sequence functions, and has become a valuable tool for functional genomics of cotton Gossypium sp. The rapid adoption of RNAi has replaced previous antisense technology.

RNAi has aided in the discovery of function and biological roles of many key cotton genes involved in fiber development, fertility and somatic embryogenesis, resistance to important biotic and abiotic stresses, and oil and seed quality improvements as well as the key agronomic traits including yield and maturity. Here, we have comparatively reviewed seminal research efforts in previously used antisense approaches and currently applied breakthrough RNAi studies in cotton, analyzing developed RNAi methodologies, achievements, limitations, and future needs in functional characterizations of cotton genes.

We also highlighted needed efforts in the development of RNAi-based cotton cultivars, and their safety and risk assessment, small and large-scale field trials, and commercialization. Today, the world demands and uses more cotton Gossypium sp.

Cotton is grown worldwide in more than 80 countries Sunilkumar et al. Uses of cotton fiber vary widely Chen et al. In addition to the cotton fiber, cottonseed products meal and hulls are used for livestock feed, and industrial lubricants Campbell et al. Cottonseed oil is used as an ingredient in food products as well as a premium cooking oil and salad dressing.

Cotton stalks and leaves are used as organic matter to enrich soil and cotton stalks have been widely used as a firewood for primary energy in cooking process in rural areas as well as conversion to biogas or composting to bio-fertilizers Federation of Indian Chambers of Commerce and Industry [FICCI] Report, Genome of allopolyploid cotton Gossypium sp.

To date, cotton lags behind many other crops in studies and gains related to genomics and genetics as well as applications of marker-assisted selection MAS due to existence of low level of molecular polymorphisms Abdurakhmonov et al. The complexity of cultivated allotetraploid cotton genomes is due to the high levels of gene redundancy because of several genome duplications events before and after allopolyploidization Adams et al.

Allopolyploidization had induced a natural gene silencing, organ specific, and homoeologously biased expression of genes regulated at developmental and epigenetic level Adams et al. Recent studies reported that a whole genome duplication WGD of ancestor-like diploid cotton Gossypium raimondii D5 had occurred approximately 60 million years ago MYA with five- to six-fold ploidy increase Renny-Byfield et al.

The gene expression analyses of this first sequenced cotton genome demonstrated complex, tissue-specific differential gene expression patterns and the diversification of gene expression level before divergence Renny-Byfield et al. The allotetraploidization of A and D genome diploids occurred about 1.

One primary objective of worldwide cotton breeding effort has been to transfer ELS fiber quality into Upland cotton while keeping their early maturity, resistance, and productivity of Upland genotypes Abdurakhmonov et al. However, traditional breeding successes have been minimal over past century of worldwide breeding, which suffered from linkage drag and distorted segregation that occurred in interspecific hybrid progenies from Upland and Pima sexual crosses Saha et al.

Besides, simultaneous improvement of fiber quality, early flowering, early maturity, and productivity in Upland cotton G. This challenge perhaps is even more severe under above-mentioned complexities of cotton genome, including higher rates of transgressive, differential and novel gene expression patterns, and homoeologous gene silencing Yoo et al. Despite this complexity, there is a need to develop Upland cotton cultivars with increased yield, early maturity, and disease and pest resistance while producing longer and stronger fibers to be competitive with synthetic fibers in the global market.

One of such innovative approaches is the expanded utilization of gene silencing techniques such as RNA interference RNAi. A survey of literature revealed that to date, RNAi has been applied for the functional studies of many agronomically, biologically, and physiologically important cotton Gossypium sp. Application of RNAi is rapidly expanding due to the advances made in cotton genomics. Efforts should be expanded further with the completed genome sequencing efforts of two ancestor-like diploid D 5 Paterson et al.

AD 1 cv. Texas Marker-1 TM-1 ; Li et al. Xinhai21; Liu et al. Here, we highlighted these efforts in detail, discussing successes, existing methods, cultivar development, field trials, safety, risks, limitations, and future perspectives of RNAi in cotton research and breeding. A Number of publications for the past years period; and B RNAi studies on the targeted cotton genes or traits.

RNAi is induced by exo- or endogenous i. RNAi is part of the normal cellular function as well as an immune response against foreign nucleic acid signatures from viral infections Roberts et al. The guide strand is incorporated into the RNA-induced silencing complex RISC , which triggers the recognition and digestion of complementary RNA sequence signatures, whereas the passenger strand gets degraded.

RISC and downstream RNAi machinery are common for exogenous and endogenous dsRNA recognition and target sequences digestion; however, there are clear differences in distinctly processing and handling of exogenous and endogenous dsRNAs Hannon, Moreover, RNAi pathway components i. At the same time, small dsRNAs may also possibly up-regulate expression of genes through binding into a promoter region and histone demethylation Li et al.

For constructing HP vectors, usually — bp long fragments of gene of interest are used. This is particularly challenging when there are several paralogous, orthologous, and highly similar gene family members in an allotetraploid genome like cultivated cotton.

These synthetic oligonucleotide-based vectors could efficiently and selectively silence target genes in plants Higuchi et al. However, both HP and synthetic oligonucleotide vectors require genetic transformation where many genotypes of cotton are recalcitrant to genetic transformation. Therefore, there is a need for rapid in vitro and in vivo testing of RNAi effects for targeted genes.

Tang et al. These efforts resulted in efficient silencing of GFP marker gene expression, which is useful for large-scale screening of gene function and drug target validation Tang et al. Due to the limitation of siRNA delivery in vitro experiments, that affect the efficiency of RNAi, researchers later developed the first efficient delivery system of siRNA to plant cells including cotton by a nanosecond pulsed laser-induced stress wave LISW.

When inoculated, such VIGS system showed efficient silencing of the target genes in tobacco, Arabidopsis and Upland cotton plants Kumar et al. A variety of Agrobacterium -mediated VIGS vectors bearing various marker genes to monitor RNAi efficiency were developed rapidly to test the gene function of interest from cotton genome. As described above and extensively referenced herein, VIGS has been widely used for the discovery and identification of many useful genes in cotton.

In this study, the gene silencing effort has resulted in a significant increase in anthocyanins and a decrease in the PAs, - -epicatechin, and - -catechin in the stems and leaves of VIGS-infected plants. It is noteworthy to mention that the first pioneering attempts on silencing of cotton genes were performed by using antisense constructs.

The pioneering effort is dated to John who studied two E6 genes isolated from Upland and Pima cottons. This research demonstrated that E6 was not critical to the fiber development process John, Antisense suppression of cotton small GTPase Rac RAC13 genes decreased the levels of H 2 O 2 in fiber cells, which in turn affected secondary wall formation of cotton fibers Potikha et al.

Ruan et al. Later, the functions of several cotton genes such as cotton steroid 5-alpha-reductase GhDET2; Luo et al. These results have suggested important roles of these genes in fiber development. Antisense technology was also utilized to regulate other aspects of cotton besides fiber genes. Inverted-repeat-based gene constructs designed for two key cotton seed-specific fatty acid desaturase genes, GhSAD-1 and GhFAD , resulted in increased levels of stearic and oleic acids in RNAi cotton lines, respectively.

Results also showed that the content of palmitic acid significantly decreased in both high-stearic and high-oleic lines, providing a promising opportunity for the development of nutritionally improved cottonseed oil Liu et al. CLCV significantly reduces boll formation and development in cotton. Antisense vectors were constructed for important genes of CLCV with the aim of affecting the virulence of this virus. Transformation of these antisense constructs into tobacco plant helped to obtain virus resistant transgenic plants Asad et al.

Gossypol is a general biocide that provides protection from insect predation, but it restricts the usefulness of cottonseed as a feed and protein source for human and monogastric animals.

In the efforts to manipulate gossypol, Martin et al. Later, Townsend et al. Similar antisense technology was also used for improvement of cottonseed oil by Sunilkumar et al. Efforts resulted in two-fold increase of the oleic acid level with an accompanying decrease of linoleic acids in transgenic cottonseeds. Further, with the discovery and understanding of the mechanisms, RNAi has been applied widely in cotton research, and it has become a major research tool for studying gene functions and breeding of novel cotton cultivars.

For the past years, more than 60 RNAi related studies have been published in cotton where we see dramatic increase of efforts after These studies have targeted functional genomics of important cotton genes including, but not limited to the fiber development, fertility and somatic embryogenesis SM , abiotic and biotic stress, cottonseed and oil quality genes of cotton Figure 1 ; Supplementary Table S1.

These efforts have resulted in the development of efficient RNAi methodologies, and RNAi-derived novel biotech cotton lines and cultivars that were subsequently utilized for the functional studies, breeding, and farming of cotton, which we have reviewed below in detail.

Cell elongation and cell cytoskeleton factors are important for fiber development. RNAi of GhACT1 significantly repressed the expression of the target and disrupted the actin cytoskeleton network in fibers, resulting in the inhibition of fiber elongation.

Researchers concluded that GhACT1 plays an important role in fiber elongation but not fiber initiation Li et al. Because cotton fibers are seed epidermal cells and resemble epidermal trichomes of leaves, researchers targeted the characterization of orthologs of plant leaf trichome initiation factors in cotton fibers. Two highly similar cotton GhMYB25 genes were characterized in allotetraploid cotton, which were acquired from A and D genome ancestors.

RNAi silencing of both GhMYB25 genes using single hairpin HP construct altered the timing of rapid fiber elongation and resulted in short fibers, dramatic reduction in trichomes on other parts of the plant, and a reduction in seed production. These results demonstrated the important functional role of MYB genes in cotton fiber and seed development Machado et al. Later, Walford et al. However, normal trichome development occurred in other parts of the cotton plant, similar to the Xu fl mutant lines.

Results of this recent report elucidated a novel mechanism of transduction of gibberellic acid signal by a homeodomain protein to promote fiber cell elongation Shan et al. Arabinogalactan proteins AGPs are involved in many aspects of plant development. Results revealed significant suppression of target genes as well as partial suppression of the expression of other FLA related genes, resulting in an inhibition of fiber initiation and fiber elongation processes.

The resulting RNAi plants had significantly shorter fibers and inferior fiber quality parameters due to disruption of the cytoskeleton network and the cellulose deposition in fiber cells Li et al. Subsequently, GhFLA1 was identified in cotton, and it was demonstrated that its suppression slowed down fiber initiation and elongation. As a consequence, plants produced significantly shorter fibers compared to the wild type controls Huang et al.

These results demonstrated the importance of cotton FLA genes in fiber length and quality improvements. GhVIN1 activity was significantly higher in fiber compared to leaf, stem, and root. It expressed at subtle level in fiberless cottonseed epidermis. Interestingly this gene started expressing in initiating fibers and reached a high-level in elongation phase, but dramatically dropped after the elongation. RNAi plants for GhVIN1 gene efficiently suppressed the transcript and negatively affected the elongation, while overexpression of GhVIN1 showed positive changes in fiber elongation rates.

It was demonstrated that GhVIN1 regulates fiber elongation in an osmotic dependent manner Wang et al. Further, RNAi- induced down-regulation of GhVIN1 expression has generated a dosage-dependent fiberless cottonseed phenotype without influencing seed development, which was due to the markedly reduction of VIN activity Wang et al. The results of Wang et al. These results demonstrated that GbPDF1 is an important genetic factor, positively conditioning cotton fiber development. The detailed characterization of GbPDF1 RNAi plants revealed a higher accumulation of hydrogen peroxide, and reduced expression of ethylene and pectin synthesis or sugar transport related genes during fiber development.

Because of their single celled-expansion-model property, developing cotton fibers have been used to elucidate cell wall formation using various transgenomic tools, including VIGS and high throughput modern experimental technologies Haigler et al. These results not only demonstrated the possibility to increase fiber length by repartitioning carbon flow but also highlighted the effectiveness of the TRV-VIGS method for rapid functional analysis of genes involved in cotton fiber development Qu et al.

RNA Interference (RNAi) Induced Gene Silencing: A Promising Approach of Hi-Tech Plant Breeding

International Journal of Medical Sciences. Journal of Cancer. Journal of Genomics. Global reach, higher impact. Journal of Genomics - Submit manuscript now Int J Biol Sci ; 10 10 RNA interference RNAi is a promising gene regulatory approach in functional genomics that has significant impact on crop improvement which permits down-regulation in gene expression with greater precise manner without affecting the expression of other genes.

RNA interference RNAi , is a powerful new technology in the discovery of genetic sequence functions, and has become a valuable tool for functional genomics of cotton Gossypium sp. The rapid adoption of RNAi has replaced previous antisense technology. RNAi has aided in the discovery of function and biological roles of many key cotton genes involved in fiber development, fertility and somatic embryogenesis, resistance to important biotic and abiotic stresses, and oil and seed quality improvements as well as the key agronomic traits including yield and maturity. Here, we have comparatively reviewed seminal research efforts in previously used antisense approaches and currently applied breakthrough RNAi studies in cotton, analyzing developed RNAi methodologies, achievements, limitations, and future needs in functional characterizations of cotton genes. We also highlighted needed efforts in the development of RNAi-based cotton cultivars, and their safety and risk assessment, small and large-scale field trials, and commercialization. Today, the world demands and uses more cotton Gossypium sp.

RNAi: What is its position in agriculture?

RNA interference and potential therapeutic applications of short interfering RNAs

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RNA interference RNAi is a powerful strategy for gene function analysis, and it is also widely studied in view of a promising use in pest control. The red palm weevil, Rhynchophorus ferrugineus Oliver Coleoptera: Curculionidae , is one of the most devastating pests of palm in the world. Conventional pest management practices are not adequate to control this insect, thus the development of efficient approaches with minimal environmental impact are needed.

Skip to search form Skip to main content You are currently offline. Some features of the site may not work correctly. DOI: Singh and S. Chaudhary and R. Sirohi and S.


ABSTRACT: RNA interference, transcriptional gene silencing, virus induced gene silencing, and of the application of this technology in woody plant.


Discovery of RNAi

RNA interference RNAi is being developed and exploited to improve plants by modifying endogenous gene expression as well as to target pest and pathogen genes both within plants i. RNAi is a natural mechanism which can be exploited to make a major contribution towards integrated pest management and sustainable agricultural strategies needed worldwide to secure current and future food production. However, there is an urgent need to develop appropriate science-based risk assessment procedures for topical RNAi applications within existing plant protection products legislation. RNAi is a natural mechanism found in most eukaryotic organisms in nature and can be exploited to improve plant health. RNAi-based technology is already being exploited, and the realized examples confirm its great potential in a range of areas of crop production and protection. Plants modified to express target dsRNAs are being assessed and regulated using existing regulatory frameworks for GMO. However, there is an urgent need to develop appropriate science-based risk assessment procedures for topical RNAi applications within existing PPP legislation.

RNAi has the potential to become a powerful therapeutic approach toward targeted and personalized medicine. The range of diseases and disorders it might address is unprecedented; from cancer to cardiovascular diseases, neurodegenerative disorders and even HIV. Even more exciting is the potential of RNAi in agriculture. RNAi has provided a way to control pests and diseases, introduce novel plant traits and increase crop yield. Using RNAi, scientists have developed novel crops such as nicotine-free tobacco, non-allergenic peanuts, decaffeinated coffee, and nutrient fortified maize among many others.

Thank you for visiting nature. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser or turn off compatibility mode in Internet Explorer. In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript. The discovery of this pathway together with the elucidation of the structure and function of short interfering RNAs — the effector molecules of RNA interference — has had an enormous impact on experimental biology. RNA interference technologies are currently the most widely utilized techniques in functional genomic studies. Furthermore, there is an intense research effort aimed at developing short interfering RNAs for therapeutic purposes.

 Три часа. Стратмор поднял брови. - Целых три часа. Так долго.

RNA Interference Technology — Applications and Limitations

Он решил подумать об этом через минуту. Сейчас ему надо было совершить давно уже откладываемую прогулку в туалетную комнату. ГЛАВА 64 Сьюзан осталась одна в тишине и сумерках Третьего узла. Стоявшая перед ней задача была проста: войти в компьютер Хейла, найти ключ и уничтожить все следы его переписки с Танкадо.

RNA Interference (RNAi) Induced Gene Silencing: A Promising Approach of Hi-Tech Plant Breeding

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2 Comments

  1. Trenwalkprerdan

    10.05.2021 at 21:16
    Reply

    PDF | RNA interference (RNAi), an evolutionarily conserved mechanism triggered by double-stranded RNA (dsRNA), causes gene silencing in.

  2. Harvey W.

    14.05.2021 at 05:38
    Reply

    RNA Interference: Biology, Mechanism, and Applications. Neema Agrawal, P. V. N. RNA silencing is a novel gene regulatory mechanism that limits the transcript level Thus, virus-induced gene silencing-based tech- niques are extremely.

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