MiRNA signatures in cell cycle sorted cells [HeLa_H295R_miRNA_seq]
ABSTRACT: For the further examination of cell cycle dependent miRNA expression profile, DNA content based fluorescence activated cell sorting (cell cycle sort) was performed on human transformed cancer cell lines. Phase-dependent miRNA expression profiling was performed on G1, S and G2 phases. Results were validated by quantitative real-time PCR. Phase-dependent miRNA expression profiling was performed on sorted human cancer (cervical - HeLa, adrenocortical - NCI-H295R) cells). G1, S and G2 phases were successfully sorted and analyzed.
Previously, drug-based synchronization procedures were used for characterizing the cell cycle dependent transcriptional program. However, these synchronization methods result in growth imbalance and alteration of the cell cycle machinery. DNA content-based fluorescence activated cell sorting (FACS) is able to sort the different cell cycle phases without perturbing the cell cycle. MiRNAs are key transcriptional regulators of the cell cycle, however, their expression dynamics during cell cycle has ...[more]
Project description:sRNA profiling from adult Taenia multiceps by Illumina high-throughput sequencing was used to increase our understanding of the molecular regulation mechanisms of the cestode,find novel biomarkers for the parasitic disease and search new strategies to control these parasites. examination of 6 individuals of adult stage of Taenia multiceps (pooled)
Project description:To understand how microRNA are involved in the complex biology of this zoonotic parasite, we describe our initial attempts to characterize small RNA in adult Dirofilaria immitis by using Illumina/Solexa deep-sequencing technology. Examination of 4 different adult Dirofilaria immitis (pooled)
Project description:We applied Illumina sequencing to identify microRNAs (miRNAs) and piwi-interacting small RNAs (piRNAs) in Dugesia japonica. Dugesia were cut up after seven day’s starvation. DJ1 is the whole Dugesia body, DJ2 is the head part, DJ3 is the tail part, DJ4 is the left part and DJ5 is the right part. Total RNA was extracted by Trizol, and preserved in ethanol, stored at -80°C until further use. 5 samples examined：DJ1,DJ2,DJ3,DJ4,DJ5. SRA study (not submitted by GEO): SRP017693
Project description:We aimed to identify miRNA regulated by alternate bearing in O. europaea. For this purpose, six olive (Olea europaea L. )(Ayvalık variety) small RNA libraries were constructed from fruits (ripe and unripe) and leaves ("on-year" and "off-year" mature -leaven in November and juvenile - leaven in July plants) and sequenced by high-throughput Illumina sequencing. Bioinformatics analyses of 93,526,915 reads identified 135 conserved miRNA, belonging to 22 miRNA families in olive tree. In addition, 38 novel miRNA were discovered in the datasets. Expression of olive tree miRNA varied greatly among the six libraries, indicating contribution of diverse miRNA in balancing between reproductive and vegetative phases. The differential expression of miRNA was evaluated on the basis of the developmental phase of the samples. Sequences of six olive miRNAs (Olea europaea L. )(Ayvalık variety) plants (ripe and unripe fruits, leaves of mature and juvenile plants of both "on-year" and "off-year") were generated by Illumina sequencing
Project description:In order to discover novel small RNAs expressed in elongated spermatids, we isolated elongated spermatids from mouse testis. The small RNA fraction (18-40nt) was cloned and sequenced from total RNA. RNAs extracted from elongated spermatids were used for high throughput sequencing analysis
Project description:In order to discover novel small RNAs expressed in mature sperm, we isolated mature sperm from mouse cauda epididymis, comparing with data from adult tesis and uterus. The small RNA fraction (18-40nt) was cloned and sequenced from total RNA of mature sperm, testis and uterus of mice. RNAs extracted from mature sperm, adult testis and uterus were used for high throughput sequencing analysis
Project description:High-throughput sequence analysis of small RNAs of TYLCV-tolerant (containing tolerance gene Ty-1) and susceptible varieties indicated that miRNA-like vsRNAs and secondary vsiRNAs are mainly contributed to hotpots. Interestingly, various characteristics of vsRNAs among each sample are consistent, but in different number of expression; Deep sequencing of degradome provided evidence for the function of vsRNAs-mediated viral transcripts slicing; And bisulfite sequencing PCR suggested the geminivirus DNA methylation induced by vsRNAs. Comparison of the expression quantity and trend of viral DNA, mRNA and vsRNA inferred that the quantity of vsRNAs is significantly corresponding to the expression level of viral mRNA. Nevertheless, vsRNAs had finite effect on inhibition of virus replication and expression. Here, we also speculated that by RDR catalysis of vsRNAs amplification, the tolerance gene Ty-1 may take an effective inhibition of viral transcription at the beginning of TYLCV infection. Examination of 6 different sRNA and 4 degradome library in 2 tomato material. The main result of our paper is distinguish the host plant sRNA and viral-derived sRNA from the .fa files (MMS_21dpi, 30dpi and TY1S_21dpi, 30dpi).
Project description:PD0325901 is involved in improving reprogramming efficiency during inducing pluripotent stem cells (iPSC) or maintain a blastocyst-like state in embryonic stem cells (ESC). However, the knowledge about this small molecule regulating miRNAs in ESC was limited. To understand the role of miRNAs during PD03-induced ESC maintenance and gain an insight how PD0325901 regulates miRNAs expression; we performed small RNA sequencing using Illumina HiSeq 2000 under the compounds treatment. The data show the miRNAs regulated by PD0325901. J1 mESCs maintained in medium containing 1000 U/mL LIF and supplemented with PD0325901 for 24 hours, J1 treated with DMSO was set as control. Then total RNA was extracted for analysis.
Project description:We report the application of illumina deep sequencing technology for high-throughput profiling of microRNA in aflatoxin B1 treated rat liver tissue, as well as normal liver tissue. miRNAs of miR-17-92 cluster, thought to be tumor activating miRNAs, were down-regulated. miR-34a, considered to be a tumor suppressor, was up-regulated.Novel miRNAs and miRNA base alteration caused by SNP or editing were predicted and summarized. Examination of 2 microRNA profiles in 2 liver tissues under different treatments.
Project description:In this study, the skin tissues were harvested from the three stages of hair follicle cycling (anagen, catagen and telogen) in a fiber-producing goat breed. In total, 63,109,004 raw reads were obtained by Solexa sequencing and 61,125,752 clean reads remained for the small RNA digitalization analysis. This resulted in the identification of 399 conserved miRNAs; among these, 326 miRNAs were expressed in all three follicular cycling stages, whereas 3, 12 and 11 miRNAs were specifically expressed in anagen, catagen, and telogen, respectively. We also identified 172 potential novel miRNAs by Mireap, 36 miRNAs were expressed in all three cycling stages, whereas 23, 29 and 44 miRNAs were specifically expressed in anagen, catagen, and telogen, respectively. Gene Ontology and KEGG pathway analyses indicated that five major biological pathways (Metabolic pathways, Pathways in cancer, MAPK signalling pathway, Endocytosis and Focal adhesion) accounting for 23.08% of target genes among 278 biological functions, indicating that these pathways are likely to play significant roles during hair cycling. the skin tissues were harvested from the three stages of hair follicle cycling (anagen, catagen and telogen) in a fiber-producing goat breed