Circular RNA Detection from High-throughput Sequencing

2017 ◽  
Author(s):  
Mohamed Chaabane ◽  
Eric C. Rouchka ◽  
Juw Won Park
Keyword(s):  
High Throughput ◽  
High Throughput Sequencing ◽  
Circular Rna ◽  
Rna Detection

scholarly journals EIF4A3-induced circular RNA PRKAR1B promotes osteosarcoma progression by miR-361-3p-mediated induction of FZD4 expression

2021 ◽  
Vol 12 (11) ◽  
Author(s):  
Zhen-hua Feng ◽  
Lin Zheng ◽  
Teng Yao ◽  
Si-yue Tao ◽  
Xiao-an Wei ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
High Throughput ◽  
Crucial Role ◽  
High Throughput Sequencing ◽  
Epithelial Mesenchymal Transition ◽  
Circular Rna ◽  
Potential Therapeutic Target ◽  
Mesenchymal Transition ◽  
Downstream Target ◽  
Underlying Mechanisms

AbstractEmerging evidence indicates that circRNAs are broadly expressed in osteosarcoma (OS) cells and play a crucial role in OS progression. Recently, cancer-specific circRNA circPRKAR1B has been identified by high-throughput sequencing and is recorded in publicly available databases. Nevertheless, the detailed functions and underlying mechanisms of circPRKAR1B in OS remains poorly understood. By functional experiments, we found that circPRKAR1B enhanced OS cell proliferation, migration, and promotes OS epithelial–mesenchymal transition (EMT). Mechanistic investigations suggested that circPRKAR1B promotes OS progression through sponging miR-361-3p to modulate the expression of FZD4. Subsequently, we identified that EIF4A3 promoted cirPRKAR1B formation through binding to the downstream target of circPRKAR1B on PRKAR1B mRNA. Further rescue study revealed that overexpression of the Wnt signalling could impair the onco-suppressor activities of the silencing of circPRKAR1B. Interestingly, further experiments indicated that circPRKAR1B is involved in the sensitivity of chemoresistance in OS. On the whole, our results demonstrated that circPRKAR1B exerted oncogenic roles in OS and suggested the circPRKAR1B/miR-361-3p/FZD4 axis plays an important role in OS progression and might be a potential therapeutic target.

scholarly journals High-throughput sequencing identified circular RNA circUBE2K mediating RhoA associated bladder cancer phenotype via regulation of miR-516b-5p/ARHGAP5 axis

2021 ◽  
Vol 12 (8) ◽  
Author(s):  
Chen Yang ◽  
Zezhong Mou ◽  
Siqi Wu ◽  
Yuxi Ou ◽  
Zheyu Zhang ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Circular Rna ◽  
Luciferase Reporter ◽  
Circular Rnas ◽  
Cell Phenotype ◽  
Invasion And Migration ◽  
Rhoa Activity

AbstractBladder cancer (BC) is known as a common and lethal urinary malignancy worldwide. Circular RNAs (circRNAs), an emerging non-coding RNA, participate in carcinogenesis process of several cancers including BC. In this study, high-throughput sequencing and RT-qPCR were applied to discover and validate abnormal high expression of circUBE2K in BC tissues. Fluorescence in situ hybridization (FISH) was used to detect hsa_circ_0009154 (circUBE2K) expression and subcellular localization in BC tissues. High circUBE2K predicted unfavorable prognoses in BCs, as well as correlated with clinical features. CCK8, transwell, EdU and wound healing assays demonstrated down-regulating circUBE2K decreased BC cell phenotype as proliferation, invasion, and migration, respectively. Further studies showed that circUBE2K promoted BC progression via sponging miR-516b-5p and enhancing ARHGAP5 expression through regulating RhoA activity. Dual-luciferase reporter, FISH and RNA pulldown assays were employed to verify the relationships among circUBE2K/miR-516b-5p/ARHGAP5/RhoA axis. Down-regulating miR-516b-5p or overexpressing ARHGAP5 restored RhoA activity mediated BC cell properties after silencing circUBE2K. Subcutaneous xenograft and metastasis model identified circUBE2K significantly increased BC cell metastasis and proliferation in-vivo. Taken together, we found that circUBE2K is a tumor-promoting circRNA in BC that functions as a ceRNA to regulate ARHGAP5 expression via sponging miR-516b-5p.

scholarly journals High-Throughput Data of Circular RNA Profiles in Human Temporal Cortex Tissue Reveals Novel Insights into Temporal Lobe Epilepsy

2018 ◽  
Vol 45 (2) ◽  
pp. 677-691 ◽  
Author(s):  
Jiaxin Li ◽  
Haijun Lin ◽  
Zhenrong Sun ◽  
Guanyi Kong ◽  
Xu Yan ◽  
...  
Keyword(s):  
Temporal Lobe Epilepsy ◽  
Temporal Lobe ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Expression Profiles ◽  
Temporal Cortex ◽  
Bioinformatic Analysis ◽  
Circular Rna ◽  
Expression Level ◽  
Circular Rnas

Background/Aims: Circular RNAs (circRNAs) are a class of long noncoding RNAs with a closed loop structure that regulate gene expression as microRNA sponges. CircRNAs are more enriched in brain tissue, but knowledge of the role of circRNAs in temporal lobe epilepsy (TLE) has remained limited. This study is the first to identify the global expression profiles and characteristics of circRNAs in human temporal cortex tissue from TLE patients. Methods: Temporal cortices were collected from 17 TLE patients and 17 non-TLE patients. Total RNA was isolated, and high-throughput sequencing was used to profile the transcriptome of dysregulated circRNAs. Quantitative PCR was performed for the validation of changed circRNAs. Results: In total, 78983 circRNAs, including 15.29% known and 84.71% novel circRNAs, were detected in this study. Intriguingly, 442 circRNAs were differentially expressed between the TLE and non-TLE groups (fold change≥2.0 and FDR≤0.05). Of these circRNAs, 188 were up-regulated, and 254 were down-regulated in the TLE patient group. Eight circRNAs were validated by real-time PCR. Remarkably, circ-EFCAB2 was intensely up-regulated, while circ-DROSHA expression was significantly lower in the TLE group than in the non-TLE group (P<0.05). Bioinformatic analysis revealed that circ-EFCAB2 binds to miR-485-5p to increase the expression level of the ion channel CLCN6, while circ-DROSHA interacts with miR-1252-5p to decrease the expression level of ATP1A2. Conclusions: The dysregulations of circRNAs may reflect the pathogenesis of TLE and circ-EFCAB2 and circ-DROSHA might be potential therapeutic targets and biomarkers in TLE patients.

scholarly journals Comprehensive circular RNA expression profile in radiation-treated HeLa cells and analysis of radioresistance-related circRNAs

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e5011 ◽  
Author(s):  
Duo Yu ◽  
Yunfeng Li ◽  
Zhihui Ming ◽  
Hongyong Wang ◽  
Zhuo Dong ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
High Throughput ◽  
Protein Interaction ◽  
Hela Cells ◽  
High Throughput Sequencing ◽  
Interaction Network ◽  
Circular Rna ◽  
Protein Protein Interaction ◽  
Hub Proteins

Background Cervical cancer is one of the most common cancers in women worldwide. Malignant tumors develop resistance mechanisms and are less sensitive to or do not respond to irradiation. With the development of high-throughput sequencing technologies, circular RNA (circRNA) has been identified in an increasing number of diseases, especially cancers. It has been reported that circRNA can compete with microRNAs (miRNAs) to change the stability or translation of target RNAs, thus regulating gene expression at the transcriptional level. However, the role of circRNAs in cervical cancer and the radioresistance mechanisms of HeLa cells are unknown. The objective of this study is to investigate the role of circRNAs in radioresistance in HeLa cells. Methods High-throughput sequencing and bioinformatics analysis of irradiated and sham-irradiated HeLa cells. The reliability of high-throughput RNA sequencing was validated using quantitative real-time polymerase chain reaction. The most significant circRNA functions and pathways were selected by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. A circRNA–miRNA–target gene interaction network was used to find circRNAs associated with radioresistance. Moreover, a protein–protein interaction network was constructed to identify radioresistance-related hub proteins. Results High-throughput sequencing allowed the identification of 16,893 circRNAs involved in the response of HeLa cells to radiation. Compared with the control group, there were 153 differentially expressed circRNAs, of which 76 were up-regulated and 77 were down-regulated. GO covered three domains: biological process (BP), cellular component (CC) and molecular function (MF). The terms assigned to the BP domain were peptidyl-tyrosine dephosphorylation and regulation of cell migration. The identified CC terms were cell–cell adherens junction, nucleoplasm and cytosol, and the identified MF terms were protein binding and protein tyrosine phosphatase activity. The top five KEGG pathways were MAPK signaling pathway, endocytosis, axon guidance, neurotrophin signaling pathway, and SNARE interactions in vesicular transport. The protein–protein interaction analysis indicated that 19 proteins might be hub proteins. Conclusions CircRNAs may play a major role in the response to radiation. These findings may improve our understanding of the role of circRNAs in radioresistance in HeLa cells and allow the development of novel therapeutic approaches.

scholarly journals Circular RNA circYPEL2: A Novel Biomarker in Cervical Cancer

Genes ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 38
Author(s):  
Xinyang Zhang ◽  
Siqi Yang ◽  
Wenbo Chen ◽  
Xin Dong ◽  
Rongyu Zhang ◽  
...  
Keyword(s):  
Cervical Cancer ◽  
Cell Lines ◽  
High Throughput ◽  
High Throughput Sequencing ◽  
Strong Stability ◽  
Circular Rna ◽  
Migration And Invasion ◽  
Circular Rnas ◽  
Cancer Hallmarks ◽  
Potential Biomarker

Cervical cancer (CC) is one of the most threatening diseases in women. Circular RNAs (circRNAs) have been reported to be cancer hallmarks, but typical circRNAs in CC were rarely indicated. Through high-throughput sequencing in CC and normal cervix tissues, circYPEL2 (hsa_circ_0005600) was proposed as a candidate circRNA. CircYPEL2 exhibited significantly high expression in CC tissue and strong stability in CC cell lines. Furthermore, knockdown and overexpression of circYPEL2 indicated the potential involvement in CC proliferation, migration and invasion. Finally, the downstream regulatory genes of circYPEL2 were investigated by knockdown experiment in CC cell lines with high-throughput sequencing. In summary, our work identified circYPEL2 as a potential biomarker for clinical research of cervical cancer.

scholarly journals Detecting Circular RNA from High-throughput Sequence Data with de Bruijn Graph

2019 ◽  
Author(s):  
Xin Li ◽  
Yufeng Wu
Keyword(s):  
High Throughput ◽  
Sequence Data ◽  
Real Data ◽  
Circular Rna ◽  
New Method ◽  
De Bruijn Graph ◽  
Circular Rnas ◽  
Real Sequence ◽  
Rna Detection ◽  
De Bruijn

AbstractCircular RNA is a type of non-coding RNA, which has a circular structure. Many circular RNAs are stable and contain exons, but are not translated into proteins. Circular RNA has important functions in gene regulation and plays an important role in some human diseases. Several biological methods, such as RNase R treatment, have been developed to identify circular RNA. Multiple bioinformatics tools have also been developed for circular RNA detection with high-throughput sequence data. In this paper, we present circDBG, a new method for circular RNA detection with de Bruijn graph. We conduct various experiments to evaluate the performance of CircDBG based on both simulated and real data. Our results show that CircDBG finds more reliable cir-cRNAs with low bias, has more efficiency in running time, and performs better in balancing accuracy and sensitivity than existing methods. As a byproduct, we also introduce a new method to classify circular RNAs based on reads alignment. Finally, we report a potential chimeric circular RNA that is found by CircDBG based on real sequence data. CircDBG can be downloaded from https://github.com/lxwgcool/CircDBG.

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