Genome Editing In R. Delemar

The dsRNA can be delivered to the worms in many ways, but in this particular experiment, the bacteria containing plasmid for dsRNA is fed to the worms. The dsRNA is able to move throughout cells in worms by the pore created by SID-1

(3) Therapeutic effect of miR15a/16 inhibitor in vivo: We will infect influenza virus with 5.0 x 10 3.375 TCID50 and then deliver the miR-15a/16 inhibitor once from 6 to 72 h later. To maximize the therapeutic effects, we will deliver the miR-15a/16 inhibitors 3 times for every 24h. Lung inflammation will be evaluated using lung histology, BAL cellularity. Alveolar macrophage activation or regulation will be evaluated using FACS analysis, cytokines, chemokines production, and viral titer.

Aim 2 is to test the hypothesis that exRNA produced by Cn during host infection of the CNS is a detectable molecular biomarker for cryptococcal infection.

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.

MiR-21 has been found to be over expressed in 20 pancreatic carcinoma tissues and cell lines compared to normal tissue or cell lines. (15)

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

Type I interferon production may be initiated in response to RNA viral infection through several RLR sensors such as RIG-I and MDA5, and inhibition or defects in these sensors has been linked to a decrease in type I interferon production.19 First, we will knockdown RIG-I and MDA5 mRNA in RAW.297 and BHK-21 cells in vitro using siRNAs specific for each. We will then analyze type I interferon production in both cell lines via flow cytometry, with BHK-21 cells acting as a control.20 Both cell lines will be infected with EEEV for 6, 12, 18, or 24 hours. We will further analyze the effect of miR-142-3p on RNA sensing by generating double knockdowns of RIG-I and miR-142-3p, or MDA5 and miR-142- 3p. miR-142-3p will be knocked down using a microRNA sponge as described earlier. As a control, whole cell lysates will be collected and subjected to western blot analysis

The main material of this experiment was HCT-116 ( human colorectal carcinoma) cells where RNA isolated. Method was applied with Rneasy Mini Kit- Qiagen. Homogenization was done by 70 % ethanol. Microcentrifuge and its tubes, pipettes, pipette tips and gloves were other materials of this experiment.

MicroRNA’s can affect the gene expression by preventing their translation onto protein. This blocking function lead to enabling the cells to adapt to change condition.

Cardiovascular disease (CVD) is the major macrovascular complication of diabetes mellitus as well as the main cause of death from clinical observation. There are about 75% of patients with type 2 diabetes have died of myocardial infarction as reported. Non-coding RNA (ncRNA) is an RNA molecule that is not translated into a protein, which include snoRNAs, microRNAs, siRNAs, piRNAs, lncRNAs and others. Recently, increasing number of evidences implicate that these ncRNAs are crucially involved in many cellular responses as well as physiological and pathological processes of several diseases. In this review, we summarize recent progress in research on functional regulatory of series of ncRNAs in cardiovascular disease with Diabetes Mellitus. Meanwhile,

Post-transcriptional gene regulation by sRNAs may occurs in various patterns of base-pairing with a target RNA resulting in different outcomes or by directly binding to proteins to modulate their function.12-14 Two classes of sRNAs are identified: trans-encoded RNAs which are transcribed from intergenic regions of the genome, and cis-encoded RNAs which are encoded on the strand complementary to coding sequences or the 5′ or 3′ untranslated region (5′ UTR, 3′ UTR) of transcripts.15-17 The family of trans-encoded sRNAs usually requires the chaperone Hfq to stabilize the often imperfect base-pairing interaction with target mRNA.18 In contrast, cis-encoded sRNAs possess a region of perfect complementarity to their target mRNA and Hfq is not needed for target binding. It shows that sRNAs are involved in many important physiological processes including anaerobic growth, nutrient availability, iron homeostasis and the response to oxidative, envelope and osmotic

This arrangement provides a convenient mechanism for the coordinated expression of a miRNA and a protein.

However, the mechanism regulating the migration of mature ecRNAs into the cytoplasm remains unclear. The linear counterparts of circRNAs, such as mRNAs or lncRNAs, can penetrate the nuclear membrane through the nuclear pore complex. Therefore, some researchers put forward the hypothesis that ecRNA export may be regulated by a mechanism similar to the regulatory mechanism of linear RNA migration [18]. The degradation pathway of circRNAs is another unanswered question. The expression level of circRNA is dynamically modulated by the balance between biogenesis and degradation of circRNAs. CircRNAs may be degraded by short interfering RNAs [13]. A recent study found that circRNAs may be cleared by extracellular vesicles or microvesicle release in mammalian cells [43]. However, this conclusion is based on an in vitro study. Whether circRNAs are degraded in vivo by a similar mechanism still needs further

Microbes are any organism that can’t be seen with the naked eye. They can be classified as bacteria, protists, fungi and sometimes even parasites. Many different types of microbes can exist within a community, which means that they have the potential to have a high species diversity (Reece et. al. 2014). Found virtually anywhere, microbes can range from being very beneficial for the environment to being