scholarly journals CPA-seq reveals small ncRNAs with methylated nucleosides and diverse termini

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Heming Wang ◽  
Rong Huang ◽  
Ling Li ◽  
Junjin Zhu ◽  
Zhihong Li ◽  
...  
Keyword(s):  
Stem Cell ◽  
High Throughput Sequencing ◽  
Noncoding Rnas ◽  
High Sensitivity ◽  
Small Noncoding Rnas ◽  
Adapter Ligation ◽  
Small Ncrnas ◽  
Mouse Tissues ◽  
Polynucleotide Kinase ◽  
Complex Landscape

AbstractHigh-throughput sequencing reveals the complex landscape of small noncoding RNAs (sRNAs). However, it is limited by requiring 5′-monophosphate and 3′-hydroxyl in RNAs for adapter ligation and hindered by methylated nucleosides that interfere with reverse transcription. Here we develop Cap-Clip acid pyrophosphatase (Cap-Clip), T4 polynucleotide kinase (PNK) and AlkB/AlkB(D135S)-facilitated small ncRNA sequencing (CPA-seq) to detect and quantify sRNAs with terminus multiplicities and nucleoside methylations. CPA-seq identified a large number of previously undetected sRNAs. Comparison of sRNAs with or without AlkB/AlkB(D135S) treatment reveals nucleoside methylations on sRNAs. Using CPA-seq, we profiled the sRNA transcriptomes (sRNomes) of nine mouse tissues and reported the extensive tissue-specific differences of sRNAs. We also observed the transition of sRNomes during hepatic reprogramming. Knockdown of mesenchymal stem cell-enriched U1-5′ snsRNA promoted hepatic reprogramming. CPA-seq is a powerful tool with high sensitivity and specificity for profiling sRNAs with methylated nucleosides and diverse termini.

scholarly journals Non-micro-short RNAs: the new kids on the block

2012 ◽  
Vol 23 (24) ◽  
pp. 4664-4667 ◽  
Author(s):  
Bijan K. Dey ◽  
Adam C. Mueller ◽  
Anindya Dutta
Keyword(s):  
High Throughput ◽  
High Throughput Sequencing ◽  
Noncoding Rnas ◽  
Mechanisms Of Action ◽  
Biological Processes ◽  
Large Group ◽  
Small Noncoding Rnas ◽  
Short Rnas ◽  
Functional Classes ◽  
Functional Relevance

The advent of ultra–high-throughput sequencing has led to the discovery of a large group of small, noncoding RNAs that are not microRNAs. The functional relevance of microRNAs has been well established over the last decade. In this Perspective, we focus on the non-micro-short RNAs that comprise a variety of functional classes and range from 16–40 nucleotides in size. We will highlight how some of these non-micro-short RNAs were discovered, as well as their biogenesis, potential mechanisms of action, and role in diverse biological processes, development, and disease. Finally, we will describe what must be done to further our understanding of these enigmatic molecules.

Small regulatory noncoding RNAs in Drosophila melanogaster: biogenesis and biological functions

2020 ◽  
Vol 19 (4) ◽  
pp. 309-323
Author(s):  
Saeed Soleimani ◽  
Zahra Valizadeh Arshad ◽  
Sharif Moradi ◽  
Ali Ahmadi ◽  
Seyed Javad Davarpanah ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Regulation Of Gene Expression ◽  
Noncoding Rnas ◽  
Critical Discussion ◽  
Rnai Pathway ◽  
Biological Functions ◽  
Small Interfering Rnas ◽  
Heterochromatin Formation ◽  
Small Noncoding Rnas ◽  
Small Ncrnas

Abstract RNA interference (RNAi) is an important phenomenon that has diverse genetic regulatory functions at the pre- and posttranscriptional levels. The major trigger for the RNAi pathway is double-stranded RNA (dsRNA). dsRNA is processed to generate various types of major small noncoding RNAs (ncRNAs) that include microRNAs (miRNAs), small interfering RNAs (siRNAs) and PIWI-interacting RNAs (piRNAs) in Drosophila melanogaster (D. melanogaster). Functionally, these small ncRNAs play critical roles in virtually all biological systems and developmental pathways. Identification and processing of dsRNAs and activation of RNAi machinery are the three major academic interests that surround RNAi research. Mechanistically, some of the important biological functions of RNAi are achieved through: (i) supporting genomic stability via degradation of foreign viral genomes; (ii) suppressing the movement of transposable elements and, most importantly, (iii) post-transcriptional regulation of gene expression by miRNAs that contribute to regulation of epigenetic modifications such as heterochromatin formation and genome imprinting. Here, we review various routes of small ncRNA biogenesis, as well as different RNAi-mediated pathways in D. melanogaster with a particular focus on signaling pathways. In addition, a critical discussion of the most relevant and latest findings that concern the significant contribution of small ncRNAs to the regulation of D. melanogaster physiology and pathophysiology is presented.

scholarly journals miRNAs in ESC differentiation

2012 ◽  
Vol 303 (8) ◽  
pp. H931-H939 ◽  
Author(s):  
Emanuele Berardi ◽  
Matthias Pues ◽  
Lieven Thorrez ◽  
Maurilio Sampaolesi
Keyword(s):  
Stem Cell ◽  
Cell Fate ◽  
Critical Role ◽  
Embryonic Stem ◽  
Noncoding Rnas ◽  
Stem Cell Differentiation ◽  
Embryonic Stem Cell Differentiation ◽  
Mammalian Development ◽  
Small Noncoding Rnas ◽  
Rna Duplexes

MicroRNAs (miRNAs) are small sequences of noncoding RNAs that regulate gene expression by two basic processes: direct degradation of mRNA and translation inhibition. miRNAs are key molecules in gene regulation for embryonic stem cells, since they are able to repress target pluripotent mRNA genes, including Oct4, Sox2, and Nanog. miRNAs are unlike other small noncoding RNAs in their biogenesis, since they derive from precursors that fold back to form a distinctive hairpin structure, whereas other classes of small RNAs are formed from longer hairpins or bimolecular RNA duplexes (siRNAs) or precursors without double-stranded character (piRNAs). An increasing amount of evidence suggests that miRNAs may have a critical role in the maintenance of the pluripotent cell state and in the regulation of early mammalian development. This review gives an overview of the current state of the art of miRNA expression and regulation in embryonic stem cell differentiation. Current insights on controlling stem cell fate toward mesodermal, endodermal and ectodermal differentiation, and cell reprogramming are also highlighted.

scholarly journals Differences in small noncoding RNAs profile between bull X and Y sperm

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9822
Author(s):  
Hao Zhou ◽  
Jiajia Liu ◽  
Wei Sun ◽  
Rui Ding ◽  
Xihe Li ◽  
...  
Keyword(s):  
Functional Analysis ◽  
Sex Differences ◽  
Embryonic Development ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Noncoding Rnas ◽  
Mature Oocyte ◽  
Small Noncoding Rnas ◽  
Nucleosomal Dna ◽  
Nucleosome Binding

The differences in small noncoding RNAs (sncRNAs), including miRNAs, piRNAs, and tRNA-derived fragments (tsRNAs), between X and Y sperm of mammals remain unclear. Here, we employed high-throughput sequencing to systematically compare the sncRNA profiles of X and Y sperm from bulls (n = 3), which may have a wider implication for the whole mammalian class. For the comparison of miRNA profiles, we found that the abundance of bta-miR-652 and bta-miR-378 were significantly higher in X sperm, while nine miRNAs, including bta-miR-204 and bta-miR-3432a, had greater abundance in Y sperm (p < 0.05). qPCR was then used to further validate their abundances. Subsequent functional analysis revealed that their targeted genes in sperm were significantly involved in nucleosome binding and nucleosomal DNA binding. In contrast, their targeted genes in mature oocyte were significantly enriched in 11 catabolic processes, indicating that these differentially abundant miRNAs may trigger a series of catabolic processes for the catabolization of different X and Y sperm components during fertilization. Furthermore, we found that X and Y sperm showed differences in piRNA clusters distributed in the genome as well as piRNA and tsRNA abundance, two tsRNAs (tRNA-Ser-AGA and tRNA-Ser-TGA) had lower abundance in X sperm than Y sperm (p < 0.05). Overall, our work describes the different sncRNA profiles of X and Y sperm in cattle and enhances our understanding of their potential roles in the regulation of sex differences in sperm and early embryonic development.

scholarly journals Conditional Hfq Association with Small Noncoding RNAs in Pseudomonas aeruginosa Revealed through Comparative UV Cross-Linking Immunoprecipitation Followed by High-Throughput Sequencing

mSystems ◽  
2019 ◽  
Vol 4 (6) ◽  
Author(s):  
Kotaro Chihara ◽  
Thorsten Bischler ◽  
Lars Barquist ◽  
Vivian A. Monzon ◽  
Naohiro Noda ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
High Throughput Sequencing ◽  
Noncoding Rnas ◽  
Cross Linking ◽  
Rna Seq ◽  
Content Type ◽  
Total Rna ◽  
Small Noncoding Rnas ◽  
Rna Chaperones

ABSTRACT Bacterial small noncoding RNAs (sRNAs) play posttranscriptional regulatory roles in cellular responses to changing environmental cues and in adaptation to harsh conditions. Generally, the RNA-binding protein Hfq helps sRNAs associate with target mRNAs to modulate their translation and to modify global RNA pools depending on physiological state. Here, a combination of in vivo UV cross-linking immunoprecipitation followed by high-throughput sequencing (CLIP-seq) and total RNA-seq showed that Hfq interacts with different regions of the Pseudomonas aeruginosa transcriptome under planktonic versus biofilm conditions. In the present approach, P. aeruginosa Hfq preferentially interacted with repeats of the AAN triplet motif at mRNA 5′ untranslated regions (UTRs) and sRNAs and U-rich sequences at rho-independent terminators. Further transcriptome analysis suggested that the association of sRNAs with Hfq is primarily a function of their expression levels, strongly supporting the notion that the pool of Hfq-associated RNAs is equilibrated by RNA concentration-driven cycling on and off Hfq. Overall, our combinatorial CLIP-seq and total RNA-seq approach highlights conditional sRNA associations with Hfq as a novel aspect of posttranscriptional regulation in P. aeruginosa. IMPORTANCE The Gram-negative bacterium P. aeruginosa is ubiquitously distributed in diverse environments and can cause severe biofilm-related infections in at-risk individuals. Although the presence of a large number of putative sRNAs and widely conserved RNA chaperones in this bacterium implies the importance of posttranscriptional regulatory networks for environmental fluctuations, limited information is available regarding the global role of RNA chaperones such as Hfq in the P. aeruginosa transcriptome, especially under different environmental conditions. Here, we characterize Hfq-dependent differences in gene expression and biological processes in two physiological states: the planktonic and biofilm forms. A combinatorial comparative CLIP-seq and total RNA-seq approach uncovered condition-dependent association of RNAs with Hfq in vivo and expands the potential direct regulatory targets of Hfq in the P. aeruginosa transcriptome.

The Prognostic and Predictive Value of microRNAs in Patients with H. pylori-positive Gastric Cancer

2019 ◽  
Vol 24 (39) ◽  
pp. 4639-4645 ◽  
Author(s):  
Seyed Mostafa Parizadeh ◽  
Reza Jafarzadeh-Esfehani ◽  
Amir Avan ◽  
Maryam Ghandehari ◽  
Fatemeh Goldani ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Tumor Suppressors ◽  
Ectopic Expression ◽  
Noncoding Rnas ◽  
World Population ◽  
Normal Flora ◽  
Small Noncoding Rnas ◽  
Control Gene Expression ◽  
The World ◽  
H Pylori

Gastric cancer (GC) has a high mortality rate with a poor 5-year survival. Helicobacter pylori (H. pylori) is present as part of the normal flora of stomach. It is found in the gastric mucosa of more than half of the world population. This bacterium is involved in developing H. pylori-induced GC due to the regulation of different micro ribonucleic acid (miRNA or miR). miRNAs are small noncoding RNAs and are recognized as prognostic biomarkers for GC that may control gene expression. miRNAs may function as tumor suppressors, or oncogenes. In this review, we evaluated studies that investigated the ectopic expression of miRNAs in the prognosis of H. pylori positive and negative GC.

scholarly journals Small Noncoding RNAs in Knee Osteoarthritis: The Role of MicroRNAs and tRNA-Derived Fragments

2021 ◽  
Vol 22 (11) ◽  
pp. 5711
Author(s):  
Julian Zacharjasz ◽  
Anna M. Mleczko ◽  
Paweł Bąkowski ◽  
Tomasz Piontek ◽  
Kamilla Bąkowska-Żywicka
Keyword(s):  
Knee Osteoarthritis ◽  
Noncoding Rnas ◽  
Pathological Process ◽  
Joint Disease ◽  
Limited Information ◽  
Genetic Changes ◽  
Small Noncoding Rnas ◽  
Knee Oa ◽  
Cartilage Homeostasis

Knee osteoarthritis (OA) is a degenerative knee joint disease that results from the breakdown of joint cartilage and underlying bone, affecting about 3.3% of the world's population. As OA is a multifactorial disease, the underlying pathological process is closely associated with genetic changes in articular cartilage and bone. Many studies have focused on the role of small noncoding RNAs in OA and identified numbers of microRNAs that play important roles in regulating bone and cartilage homeostasis. The connection between other types of small noncoding RNAs, especially tRNA-derived fragments and knee osteoarthritis is still elusive. The observation that there is limited information about small RNAs different than miRNAs in knee OA was very surprising to us, especially given the fact that tRNA fragments are known to participate in a plethora of human diseases and a portion of them are even more abundant than miRNAs. Inspired by these findings, in this review we have summarized the possible involvement of microRNAs and tRNA-derived fragments in the pathology of knee osteoarthritis.

scholarly journals Development of a capture sequencing assay for enhanced detection and genotyping of tick-borne pathogens

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Komal Jain ◽  
Teresa Tagliafierro ◽  
Adriana Marques ◽  
Santiago Sanchez-Vicente ◽  
Alper Gokden ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Sequencing ◽  
Fold Increase ◽  
The United States ◽  
Genomic Research ◽  
Babesia Microti ◽  
Hybridization Capture ◽  
Mouse Tissues ◽  
Genome Analyses ◽  
Capture Sequencing

AbstractInadequate sensitivity has been the primary limitation for implementing high-throughput sequencing for studies of tick-borne agents. Here we describe the development of TBDCapSeq, a sequencing assay that uses hybridization capture probes that cover the complete genomes of the eleven most common tick-borne agents found in the United States. The probes are used for solution-based capture and enrichment of pathogen nucleic acid followed by high-throughput sequencing. We evaluated the performance of TBDCapSeq to surveil samples that included human whole blood, mouse tissues, and field-collected ticks. For Borrelia burgdorferi and Babesia microti, the sensitivity of TBDCapSeq was comparable and occasionally exceeded the performance of agent-specific quantitative PCR and resulted in 25 to > 10,000-fold increase in pathogen reads when compared to standard unbiased sequencing. TBDCapSeq also enabled genome analyses directly within vertebrate and tick hosts. The implementation of TBDCapSeq could have major impact in studies of tick-borne pathogens by improving detection and facilitating genomic research that was previously unachievable with standard sequencing approaches.

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