Assessment of different enrichment methods revealed the optimal approach to identify bovine circRNAs
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ABSTRACT:
PROVIDER: PRJNA941278 | ENA |
REPOSITORIES: ENA
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Project description:To better study the bovine circRNA, we assessed the outcomes of circRNA identification using six enrichment approaches. RNA-sequencing analysis revealed that different approaches led to varied the ratio of uniquely mapped reads, false positive rate of identifying circRNAs, and the number of circRNAs per million clean reads. Besides, 507 of 4,051 identified bovine high confident circRNAs had shared splicing sites with human circRNAs.
2024-06-05 | GSE226717 | GEO
Project description:Small RNA sequencing across diverse biofluids identifies optimal methods for exRNA isolation
| PRJNA671225 | ENA
Project description:Small RNA sequencing across diverse biofluids identifies optimal methods for exRNA isolation
| PRJNA671227 | ENA
Project description:Methylome-wide association studies (MWAS) are typically performed using microarray technologies that assay only a very small fraction of the CG methylome and entirely miss two forms of methylation that are common in brain and likely of particular relevance for neuroscience and psychiatric disorders. The alternative is the use whole genome bisulfite sequencing, but this approach is not yet practically feasible with the sample sizes required for adequate statistical power. We argue for revisiting methylation enrichment methods that, provided optimal protocols are used, enable comprehensive, adequately powered, and cost-effective genome-wide investigations of the methylome. To support our claim we use data showing that enrichment methods approximate the sensitivity obtained with the whole genome bisulfite methods with slightly better specificity. However, this performance is achieved at <5% of the reagent costs. Furthermore, because many more samples can be sequenced simultaneously, projects can be completed about 15 times faster. A potential drawback of our approach to assay CG methylation (the dominant form of methylation in the vast majority of somatic tissues). is the relatively large amount of genomic DNA (ideally >1 ug) required to obtain high quality data. Biomaterials are typically expensive to collect, provide a finite amount of DNA, and may simply not yield sufficient starting material. Thus, any possibility to use low amounts of DNA will increase the breadth and number of the studies that can be conducted. Therefore, we further adapted the enrichment step of the protocol. With this low starting material protocol, the assay performed generally equal well or better than the ample starting material protocol requiring >1 ug.
2017-02-15 | GSE94866 | GEO
Project description:To explore the overall circRNAs involved in growth and development of Arabidopsis thaliana across the lifespan, we deeply sequenced samples of whole plants from different developmental stages (cotyledons emergence, rosette leavesï¹¥1 mm, rosette growth complete, first flower open, flourishing florescence, first silique shattered, senescence). The total RNA was purified by rRNA-depletion and linear RNA removal with RNAseR, and sequenced by the Illumina HiSeq2500 platform. We obtained 31 Gb raw data and identified 1217 circRNAs with expression quantity. We annotated these circRNAs and predicted their targeted microRNA. The circRNAs involved in growth and development of Arabidopsis thaliana across lifespan were identified and analyzed using the Illumina HiSeq2500 platform.
2016-09-01 | E-GEOD-77672 | biostudies-arrayexpress
Project description:Objective: As one kind of novel non-coding RNA with high stability and conservatism, circular RNAs (circRNAs) are involved in different biological processes. Although growing evidence has supported the important role of circRNAs in renal diseases, the specific mechanism still remains unknown in neonatal acute kidney injury (AKI). Methods: High throughput sequencing analysis was applied to investigate the expression patterns of circRNAs in the comparison between hypoxia induced AKI neonates and controls. Then, bioinformatics analysis was conducted to predict the function of differentially expressed circRNAs and construct the network of circRNA-miRNA interaction. Results: (1) We identified 296 differentially expressed circRNAs (circRNAs with Fold change > 2 and P value < 0.05). Of them, 184 circRNAs were markedly up-regulated and 112 were significantly down-regulated in the AKI group. (2) Bioinformatics analysis indicated that the parental genes of the differentially expressed circRNAs were predominantly implicated in the cell and cell part, cellular process and cancer pathways. Conclusion: This study could contribute to future research of circRNAs in the pathogenesis of neonatal AKI and facilitate the identification of novel therapeutic targets of circRNAs.
2023-01-11 | GSE222578 | GEO
Project description:Background: Ischemic stroke is a disease with high rate of death and disability worldwide. This study investigated key circRNAs related to ischemic stroke. Methods: Three ischemic stroke patients and three healthy individuals were included in the current study to obtain the circRNA expression profiles by RNA sequencing. Through bioinformatic analysis, differentially expressed circRNAs (DEcircRNAs) were identified, and GO and pathway analyses for the host genes of DEcircRNAs were conducted. To further explore the functions of key circRNAs, a DEcircRNA-miRNA interaction network was constructed. Finally, the expression levels of selected circRNAs were validated with qRT-PCR. Results: A total of 736 DEcircRNAs were detected in ischemic stroke. Functional annotation of host genes of DEcircRNAs revealed several significantly enriched pathways, including Fc epsilon RI signaling pathway, B cell receptor signaling pathway, and T cell receptor signaling pathway. A circRNA-miRNA network, including 1544 circRNA-miRNA pairs, 456 circRNAs and 4 miRNAs, was obtained. The qRT-PCR results were largely in keeping with our RNA-seq data. Conclusion: The results of our study may help to elucidate the specific mechanism underlying ischemic stroke.
2021-06-24 | GSE178764 | GEO
Project description:Purpose: With the advent of human genome sequencing project, circRNAs attracted widespread attention in cancer research due to its stable ring structure. Our aim was to identify differentially expressed circRNAs in GC and explore their potential roles in GC diagnosis and treatment. Methods: Total RNA from tissues was isolated by Hipure Total RNA Mini Kit (Magen, Germany). Qubit 3.0 Fluorometer (Invitrogen, Carlsbad, California) was used for RNA concentration measurement while Agilent 2100 Bioanalyzer (Applied Biosystems, Carlsbad, CA) for RNA integrity estimation. A RIN value over 7.0 was considered eligible. RNA-seq library was prepared with approximately 2μg of total RNA using KAPA RNA HyperPrep Kit with RiboErase (HMR) for Illumina® (Kapa Biosystems, Inc., Woburn, MA). Briefly, total RNA was incubated at 37 °C for 30 min with 10 units RNase R (Epicentre Technologies, Madison, USA) after removing ribosomal RNA. Next, the ribominus RNase R (+) RNAs was fragmented and then first strand and directional second strand synthesis were performed. Then the A tailing and adapter ligation were performed with the purified cDNA. Finally, the purified, adapter-ligated DNA was amplified. Each library was diluted to 10 nM and pooled equimolar prior to clustering. Paired-End (PE150) sequencing was performed on all samples. Results: A total of 25,303 circRNA targets, including 20,036 known circRNAs and 5,267 uncharacterized circRNAs, were detected and defined. Among them, there were 2,007 circRNAs with statistically significant differences in expression in GC tissues, in which fold changes>2.0 and P<0.05 were identified.
2019-06-16 | GSE131414 | GEO
Project description:Intensive research in past two decades has uncovered the presence and importance of noncoding RNAs (ncRNAs), which includes microRNAs (miRs) and long ncRNAs (lncRNAs). These two classes of ncRNAs interact to a certain extent, as some lncRNAs bind to miRs to sequester them. Such lncRNAs are collectively called 'competing endogenous RNAs' or 'miRNA sponges'. In this study, we screened for lncRNAs that may act as miRNA sponges using the publicly available data sets and databases. To uncover the roles of miRNA sponges, loss-of-function experiments were conducted, which revealed the biological roles as miRNA sponges. LINC00324 is important for the cell survival by binding to miR-615-5p leading to the de-repression of its target BTG2 LOC400043 controls several biological functions via sequestering miR-28-3p and miR-96-5p, thereby changing the expressions of transcriptional regulators. Finally, we also screened for circular RNAs (circRNAs) that may function as miRNA sponges. The results were negative at least for the selected circRNAs in this study. In conclusion, miRNA sponges can be identified by applying a series of bioinformatics techniques and validated with biological experiments. The expressions of miRs were examined in HEK-293 cells that were LINC00324- or LOC400043-silenced using siRNA.
2016-07-06 | E-GEOD-73689 | biostudies-arrayexpress
Project description:Intensive research in past two decades has uncovered the presence and importance of noncoding RNAs (ncRNAs), which includes microRNAs (miRs) and long ncRNAs (lncRNAs). These two classes of ncRNAs interact to a certain extent, as some lncRNAs bind to miRs to sequester them. Such lncRNAs are collectively called 'competing endogenous RNAs' or 'miRNA sponges'. In this study, we screened for lncRNAs that may act as miRNA sponges using the publicly available data sets and databases. To uncover the roles of miRNA sponges, loss-of-function experiments were conducted, which revealed the biological roles as miRNA sponges. LINC00324 is important for the cell survival by binding to miR-615-5p leading to the de-repression of its target BTG2 LOC400043 controls several biological functions via sequestering miR-28-3p and miR-96-5p, thereby changing the expressions of transcriptional regulators. Finally, we also screened for circular RNAs (circRNAs) that may function as miRNA sponges. The results were negative at least for the selected circRNAs in this study. In conclusion, miRNA sponges can be identified by applying a series of bioinformatics techniques and validated with biological experiments. The expressions of miRs were examined in HEK-293, HUVEC and Hs68 cells.
2016-07-06 | E-GEOD-73690 | biostudies-arrayexpress