The Role Of ERNA And Exrna

The goal of this project is to determine the effect of the RNAi targeting the gene unc-22 in C. Elegans through visual observation of worm

MicroRNAs (miRNA) are small noncoding RNA, usually 17-25 nucleotides long that are able to bind complementary sequences of target messenger RNA (mRNA) and to induce both their degradation and translational repression (Fortunato, et al 2014). They are one of the most significant classes of non-coding RNA molecules (eg. small interfering RNA (siRNA) and ribozymes) that act within the cell. MiRNAs are also evolutionary conserved in different species from plants to humans and are encoded by their respective genes (Bader, 2012).

qRT-PCR was used in Figure 1 to confirm that the transfection with siRNA was successful and evaluate the siRNA knockdown of CDK8 in RNA.

In the second paper, “Novel RNA Modifications in the Nervous System: Form and Function,” Lee discusses the role of several novel RNA modification in the physiological setting of the nervous system. RNA editing plays a major role in many pathologies; for example, loss of normal function in an m6a demethylase, FTO, has been connected with cancer, ADHD, and Alzheimer’s. Other RNA modifications are necessary for proper development of the central nervous system. NSUN2 is an m5C RNA modification heavily implicated in embryogenesis, especially in the brain. Mutation of the NSUN2 gene is correlated with intellectual disability, developmental delay, and facial dysmorphism. The aforementioned A-to-I RNA edit is also enriched in the brain, and is necessary for the proper function of genes such as GluA2, which without modification leads to postnatal death. GluA2 is also implicated in glutamate reception by neurons, and consequently in the mechanisms of addiction. Furthermore, A-to-I RNA editing failure been linked with several chronic diseases, including

This study shows that 6% of the entire annotated non-coding and coding gene transcripts are overlapping with small RNAs. Highly specific subcellular positioning is found for both unannotated & annotated short RNA.

Rhizopus delemar is an emerging fungal pathogen that causes devastating mucormycosis in individuals with a weakened immune system. The symptoms of mucormycosis are non-specific and definitive diagnosis requires time-consuming and often insensitive direct examination of specimens and cultures of the organisms. Recently, extracellular vesicles (EVs) with encapsulated RNAs (exRNAs), have emerged as an important cell-cell communication conduit that holds great promise not only in diagnoses but also treatments of cancer and infectious diseases. We have performed deep-sequencing, profiling and annotation of extracellular small non-coding ribonucleic acids (ex-sRNAs) isolated from two clinical strains of R. delemar. Approximately 3.3 and 3.2

Lab Report 3: Identification of soil organism by sequencing 16S RNA and characterization of Streptomyces

The quantity of each product is then analyzed using qRT-PCR and the brains characterized to see if adding these genes has rescued the function of ADCK3 in the RNAi fly line

The intriguing finding that Cin1-S confers a CNS survival advantage provides a novel avenue to probe the CNS propensity of Cn. Finally, given the importance of exRNA in intercellular cellular communication, it is tempting to speculate that Cin1 regulates uptake and export of exRNA to play a role in host-pathogen interactions and pathogenesis. Because of our recent advances, we believe we may possess an unprecedented system to probe the mechanism of pathogenesis in Cn.

There is no mentioning alternative and more sensitive test (Reviewer 2). The reviewer commented that we might miss mild effects of lncRNAs in plasticity and memory storage. Mild effects of lncRNA expression changes will be difficult to dissect in vivo. Given that lncRNAs are critical mediators of transcription and translation, the two key processes that regulate long-term synaptic plasticity, we are confident that we will be able to identify lncRNAs that are critical for synaptic plasticity and memory. We are certainly interested in the mild effects of lncRNAs, but during the two-year funding of this project, we sought to identify and characterize lncRNAs that are necessary for synaptic plasticity and memory

It is triggered by Dicer, an enzyme that assists in the activation of the RNA induced silencing complex, an important component for RNAi in fungi, plants, and animals, according to the article, “Pathway Central: RNAi Pathway”. The process is first activated by dsRNA molecules and requires a specific set of gene products. The dsRNA is then cut into smaller pieces by the Dicer enzyme in an ATP-dependent reaction. The dsRNA in fungi, animals, and plants are exogenous, meaning that the RNA is directly sent to the cytoplasm and cut into shorter lengths by Dicer. As of yet, endogenously expressed dsRNA molecules have not been discovered in mammals, where the RNA would first be shaped into the stem- loop structure in the nucleus, then sent to the cytoplasm to be further modified by Dicer. In RISC, the enzyme Dicer also supplies the initial RNA material to activate the complex as well as the first RNA substrate molecule. RISC with a bound siRNA targets complementary mRNA molecules and degrades them, resulting in lower levels of protein translation and the eventual inhibition of the

MiR-142-3p and mir-142-5p were depleted in Ago-IP profile, compared to total cellular profile, at least two-fold within each cell lines, separately.

short RNA. One of these strands of RNA contains the matching sequence to that of the attacking

We will perform expression validation of exRNA signatures produced by JEC21, a serotype D strain and analyze whether these exRNA signatures are also present in other serotype D strains of Cn, as well as serotype B and C C. gattii strains using RT-qPCR. We will isolate exRNAs from serotype A Cn strain H99 and Cg strains such as R265 (VGIIa) and R272 (VGIIb). The results will be analyzed to look for common expression profiles of RNA that could serve as molecular signatures produced by Cn and possibly Cg strains that are not produced by mammalian hosts such as humans or mice. This bioinformatics work will be performed under the direction of the in-house bioinformatics specialist, Dr. Chris Taylor. Please see his attached letter of support. Production of the biomarker candidates will be verified by RT-qPCR.

The lncRNAs transcripts will be given by the defined reference noncoding genome (Aim 1.1). An effect size of differential lncRNA transcript methylation (M_i) will be calculated for reference region i, similar as in Formula 2.