Genome-wide computational prediction and experimental validation of quinoa (Chenopodium quinoa) microRNAs

2019 ◽  
Vol 99 (5) ◽  
pp. 666-675 ◽  
Author(s):  
Ashutosh Sharma ◽  
Paola Isabel Angulo Bejerano ◽  
Irais Castillo Maldonado ◽  
Marcos de Donato Capote ◽  
Alfredo Madariaga-Navarrete ◽  
...  

MicroRNAs (miRNAs) are highly conserved, endogenous, short (21–24 nucleotides), non-coding RNA molecules that play significant roles in post-transcriptional gene silencing by directing target mRNA cleavage or translational inhibition. Nonetheless, highly nutritious “super grain” quinoa (Chenopodium quinoa) is an extreme abiotic stress tolerant Andean seed crop of many potential uses, with outstanding protein quality and a load of vitamins, minerals, as well as flavonoid antioxidants. In this study, applying genome-wide in silico approaches (referring to the recently published quinoa genome) and following a set of stringent filtering measures, a total of 22 potentially conserved microRNAs belonging to 18 families were characterized from quinoa and 11 randomly selected putative microRNAs (cqu-miR160a, cqu-miR162a, cqu-miR164a, cqu-miR166b, cqu-miR167a, cqu-miR172a, cqu-miR319a, cqu-miR390a, cqu-miR393a, cqu-miR394a, and cqu-miR398b) were validated successfully by RT-PCR. Using the psRNATarget tool, a sum of 59 potential miRNA targets, mostly transcription factors, were identified that are involved in biosynthesis, metabolic processes, and signal transduction. Among the detected targets, six target transcripts (F-Box proteins, TCP, MYB, WD protein, NAC, and CSD) were reported to have specific roles in both flavonoids biosynthesis and stress response signaling in some plants. To the best of our knowledge, this is the first report of quinoa microRNAs and their targets.

Zygote ◽  
2019 ◽  
Vol 27 (05) ◽  
pp. 263-271 ◽  
Author(s):  
Maryam Kiani ◽  
Mohammad Salehi ◽  
Asghar Mogheiseh

SummaryInfertility is an important reproductive health problem, and male infertility is especially important in more than half of infertility cases. Due to the importance of genetic factors in this condition, analysis of semen alone is not enough to recognize men with idiopathic infertility. A molecular non-invasive investigation is necessary to gain valuable information. Currently, microRNAs (miRNAs) are being used as non-invasive diagnostic biomarkers. miRNAs, single-stranded non-coding RNA molecules, act as post-transcriptional gene silencing regulators either by inhibition or repression of translation. Changes in the regulation of miRNAs have been investigated in several different types of male infertility, therefore the biological role of miRNA and gene targets has been defined. The purpose of this study was to review recent research on the altered expression of miRNA in semen, sperm, and testicular biopsy samples in infertile males with different types of unexplained infertility. Changes in miRNA regulation were investigated using microarray and the miRNA levels were confirmed by real-time qRT-PCR. This review explains why creating a non-invasive diagnostic method for male infertility is necessary and how changes in miRNA expression can be used as new diagnostic biomarkers in patients with differing spermatogenic and histopathologic injury.


2021 ◽  
Author(s):  
Wittaya Chaiwangyen

MicroRNAs (miRNAs) are a class of non-coding endogenous RNA molecules that are involved in post-transcriptional gene silencing via binding to their target messenger RNA, leading to mRNA degradation or translational repression. MicroRNAs can be modulated by several factors including hormones, transcription factors, and dietary compounds. These biologically active compounds have positive impact on the progression of human pathology including non-communicable diseases, which indicating that administration of diet may have potential as therapeutic agents in modulating the risk of chronic diseases. Interestingly, evidence emerging in recent years suggests that dietary miRNAs can be absorbed in human circulation, modulated human gene expression and biological functions. The exploitation of the miRNA functioning within different origins, cellular miRNAs and dietary miRNAs will help us to understand the molecular machinery as well as the regulatory mechanisms involved in fundamentally important biological processes. Therefore, this knowledge may be applied of natural bioactive compounds in preventive or therapeutic approaches.


Author(s):  
Angela Ricci ◽  
Silvia Sabbadini ◽  
Laura Miozzi ◽  
Bruno Mezzetti ◽  
Emanuela Noris

Abstract Since the beginning of agriculture, plant virus diseases have been a strong challenge for farming. Following its discovery at the very beginning of the 1990s, the RNA interference (RNAi) mechanism has been widely studied and exploited as an integrative tool to obtain resistance to viruses in several plant species, with high target-sequence specificity. In this chapter, we describe and review the major aspects of host-induced gene silencing (HIGS), as one of the possible plant defence methods, using genetic engineering techniques. In particular, we focus our attention on the use of RNAi-based gene constructs to introduce stable resistance in host plants against viral diseases, by triggering post-transcriptional gene silencing (PTGS). Recently, spray-induced gene silencing (SIGS), consisting of the topical application of small RNA molecules to plants, has been explored as an alternative tool to the stable integration of RNAi-based gene constructs in plants. SIGS has great and innovative potential for crop defence against different plant pathogens and pests and is expected to raise less public and political concern, as it does not alter the genetic structure of the plant.


2020 ◽  
Vol 21 (10) ◽  
pp. 3711
Author(s):  
Melina J. Sedano ◽  
Alana L. Harrison ◽  
Mina Zilaie ◽  
Chandrima Das ◽  
Ramesh Choudhari ◽  
...  

Genome-wide RNA sequencing has shown that only a small fraction of the human genome is transcribed into protein-coding mRNAs. While once thought to be “junk” DNA, recent findings indicate that the rest of the genome encodes many types of non-coding RNA molecules with a myriad of functions still being determined. Among the non-coding RNAs, long non-coding RNAs (lncRNA) and enhancer RNAs (eRNA) are found to be most copious. While their exact biological functions and mechanisms of action are currently unknown, technologies such as next-generation RNA sequencing (RNA-seq) and global nuclear run-on sequencing (GRO-seq) have begun deciphering their expression patterns and biological significance. In addition to their identification, it has been shown that the expression of long non-coding RNAs and enhancer RNAs can vary due to spatial, temporal, developmental, or hormonal variations. In this review, we explore newly reported information on estrogen-regulated eRNAs and lncRNAs and their associated biological functions to help outline their markedly prominent roles in estrogen-dependent signaling.


HortScience ◽  
2005 ◽  
Vol 40 (4) ◽  
pp. 1050C-1050
Author(s):  
Kullanart Obsuwan ◽  
Wayne B. Borth ◽  
John Hu ◽  
Adelheid R. Kuehnle

A Cymbidium mosaic virus movement protein gene with a site-specific mutation (mut11) under control of a ubiquitin promoter was inserted using biolistics into two Dendrobium varieties with the intention of creating CymMV-resistant orchids. Presence of the transgene in regenerated plants of D. × Jaquelyn Thomas `Uniwai Mist' and D. x Jaq–Hawaii `Uniwai Pearl' was confirmed by PCR using genomic DNA, and mut11-positive plants were potted ex vitro. Forty-two transgenic plants and four non-transgenic control plants at the 4- to 6-leaf stage were inoculated with a 1:1000 dilution of CymMV obtained from infected orchids. Plants were analyzed for systemic infection using tissue blot immunoassay (TBIA). Seventeen plants from at least six independent transformations remained virus-free, whereas all control plants were infected with CymMV within 1 month. Further analysis by RT-PCR showed that the mut-11 mRNA was detectable in only two of these 17 plants. All plants were challenged again with a second CymMV inoculation as above, followed by TBIA analysis after 1 month. Thirteen of 17 plants remained free from virus. A third challenge of these plants with CymMV as above was followed by TBIA analysis at 1 week, 2 weeks, 1 month, 3 months, 6 months, and 12 months after challenge. Results at 2 weeks post-inoculation showed that all six controls and four individual transgenic plants, including the RT-PCR-positive plants, became systemically infected. Nine transgenic plants from both varieties remained free from CymMV 12 months after the third challenge. Lack of detectable mut11 mRNA in these resistant lines suggests that a post-transcriptional gene silencing (PTGS) mechanism may be conferring resistance to CymMV.


2018 ◽  
Author(s):  
Marius van den Beek ◽  
Bruno da Silva ◽  
Juliette Pouch ◽  
Mohammed el amine Ali Chaouche ◽  
Clément Carré ◽  
...  

AbstractpiRNA-mediated repression of transposable elements (TE) in the germline limits the accumulation of heritable mutations caused by their transposition in the genome. It is not clear whether the piRNA pathway plays a functional role in adult, non-gonadal tissues in Drosophila melanogaster. To address this question, we first analyzed the small RNA content of adult Drosophila melanogaster heads. We found that varying amount of piRNA-sized, ping-pong positive molecules in heads correlates with contamination by gonadal tissue during RNA extraction, suggesting that most of piRNAs detected in head sequencing libraries originate from gonads. We next sequenced the heads of wild type and piwi mutants to address whether piwi loss of function would affect the low amount of piRNA-sized, ping-pong negative molecules that are still detected in heads hand-checked to avoid gonadal contamination. We find that loss of piwi does not affect significantly these 24-28 RNA molecules. Instead, we observe increased siRNA levels against the majority of Drosophila transposable element families. To determine the effect of this siRNA level change on transposon expression, we sequenced the transcriptome of wild type, piwi, dicer-2 and piwi, dicer-2 double-mutant fly heads. We find that RNA expression levels of the majority of TE families in piwi or dicer-2 mutants remain unchanged and that TE transcript abundance increases significantly only in piwi, dicer-2 double-mutants. These results lead us to suggest a dual-layer model for TE repression in adult somatic tissues. Piwi-mediated transcriptional gene silencing (TGS) established during embryogenesis constitutes the first layer of TE repression whereas Dicer-2-dependent siRNA-mediated post-transcriptional gene silencing (PTGS) provide a backup mechanism to repress TEs that escape silencing by piwi-mediated TGS.


2004 ◽  
Vol 82 (4) ◽  
pp. 472-481 ◽  
Author(s):  
Tony Nolan ◽  
Carlo Cogoni

Small RNA molecules such as siRNAs and miRNAs represent a new class of molecules that have been implicated in a wide range of diverse gene silencing phenomena. It is now becoming clear that these two similar molecules share several common features in both their biogenesis and their mechanism of action. Thus, the siRNA and miRNA pathways may have evolved from a common ancestral mechanism that has diverged to play important roles in developmental regulation, genomic organisation, and cellular defence against foreign nucleic acids.Key words: miRNA, siRNA, post-transcriptional gene silencing, RNAi, heterochromatin.


2005 ◽  
Vol 386 (12) ◽  
pp. 1219-1238 ◽  
Author(s):  
Jörg Vogel ◽  
Cynthia Mira Sharma

AbstractSmall non-coding RNAs (sRNAs) have attracted considerable attention as an emerging class of gene expression regulators. In bacteria, a few regulatory RNA molecules have long been known, but the extent of their role in the cell was not fully appreciated until the recent discovery of hundreds of potential sRNA genes in the bacteriumEscherichia coli. Orthologs of theseE. colisRNA genes, as well as unrelated sRNAs, were also found in other bacteria. Here we review the disparate experimental approaches used over the years to identify sRNA molecules and their genes in prokaryotes. These include genome-wide searches based on the biocomputational prediction of non-coding RNA genes, global detection of non-coding transcripts using microarrays, and shotgun cloning of small RNAs (RNomics). Other sRNAs were found by either co-purification with RNA-binding proteins, such as Hfq or CsrA/RsmA, or classical cloning of abundant small RNAs after size fractionation in polyacrylamide gels. In addition, bacterial genetics offers powerful tools that aid in the search for sRNAs that may play a critical role in the regulatory circuit of interest, for example, the response to stress or the adaptation to a change in nutrient availability. Many of the techniques discussed here have also been successfully applied to the discovery of eukaryotic and archaeal sRNAs.


2010 ◽  
Author(s):  
Krishnan Namboori ◽  
Arun Mohan ◽  
Radhagayathri K.U. ◽  
Varun Gopal ◽  
Vasavi C.S. ◽  
...  

3 Biotech ◽  
2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Sujay Paul ◽  
Juan Luis de la Fuente-Jiménez ◽  
Camila Garibay Manriquez ◽  
Ashutosh Sharma

AbstractmicroRNAs (miRNAs) are highly conserved, short (~ 21-nucleotide), endogenous, non-coding RNA molecules that play major roles in post-transcriptional silencing by guiding target mRNA cleavage or translational inhibition. In this study, applying high-stringent genome-wide computational-based approaches, a total of 28 putative miRNAs belonging to 17 miRNA families were identified from an antioxidant-rich medicinal plant passion fruit (Passiflora edulis). Inter-tissue (leaves and fruits) and inter-varietal (yellow and purple fruit varieties) quantitative study of six putative passion fruit miRNAs (ped-miR160, ped-miR164, ped-miR166, ped-miR393, ped-miR394, and ped-miR398) showed differential expression. Using psRNATarget tool, a total of 25 potential target proteins of the characterized passion fruit miRNAs were also identified. Most of the target proteins identified in this study, including SQUAMOSA promoter binding, Class III HD-Zip, NAC, Scarecrow, APETALA2, Auxin response factor, MYB, and superoxide dismutase, were found to be involved in development, metabolism, and defense/stress response signaling.


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