taking a Look at MicroRNAs

In this project, C. Elegans are hermaphrodite worms that will be used since they are easy to maintain in lab, as well as have short life cycles. The gene that the project attempted to knockdown in C. Elegans with RNAi treatment is the unc-22 gene. RNAi disrupts gene expression in the presence of double stranded RNA (dsRNA) that is complementary to target gene sequence. The unc-22 gene codes for a muscle protein called twitchin in wild-type worms. The Unc-22 is required for muscle regulation and maintenance in C.Elegans. To verify that the RNAi treatment worked, would check the unc-22 mRNA levels in the worms, in addition to phenotype observation.

An abnormality in this process leads to a number of problems, including cancer. MicroRNAs are a type of RNA which inhibit the expression of genes and play a vital role in the development of cancer. Of notable interest is miR-378, as it increases tumor angiogenesis and growth and also enhances the survival of these cells, by decreasing the expression of tumor suppression genes like Sufu. A study testing human cell lines found that when Sufu expression levels were high, miR-378 expression levels were low, and when Sufu levels were low, miR-378 levels were high, showcasing that the two were negatively correlated. Because repression of Sufu can lead to rampant proliferation of cells, the inhibition of Sufu is essential in order for miR-378 to promote angiogenesis and increase cell

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).

Recent reports indicate the potential role of miRNAs as both therapeutic targets and biomarkers in IPF (5). MiRNAs are small (approximately 22 nucleotides in length) and stable non-coding molecules, which can regulate gene expression (6). They are important regulators of physiological processes, such as cellular proliferation leading to lung tissue repair. The miR200 family is functionally involved in canonical pathways of immune response and cell-cell communication. Here we propose novel studies using miR200b and miR200c in IPF compared to controls, first, to determine their function in the niche composed of ATII cells and fibroblasts, second, analyze their potential to stimulate ATII cell proliferation and transdifferentation to alveolar type I cells, third to analyze their capacity to inhibit the growth of fibroblasts; and fourth, to use them as early biomarkers of IPF development. The rationale for using 200miRNA family members as curing IPF drug is based on their dysregulated expression observed in IPF (7). Moreover, our

The RNA-induced-silencing complex is used for gene regulation and in defense against viral and bacterial infections. The RISC is loaded with small dsRNAs that will guide the complex through base-pairing interactions to specific messenger RNA targets, which the RISC will help degrade. Once degraded, no more protein can be made from these mRNAs. The RNA-induced silencing complex can silence targeted genes through several mechanisms. It can work to repress translation during protein synthesis, interfere with the transcript level through mRNA degradation, or work at the level of the genome itself and eliminate DNA/form degrading materials. Heterochromatin is a degrading material that can be created to modify or suppress genes in the RISC (MacRae

1. Do you think that sexual orientation is Myrna’s primary counseling issue? Explain your rationale.

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

We have also noticed, the absence of miR-4792, miR-4485-3p, miR-4532 and miR-486-5p in our Ago2-complex, compared to total cellular miRNA profile. No proper signal was detected, using NB. These miRNAs might be part of transfer RNA, or associated with other type of Ago protein, or could be a degraded tRNA product. The function of aforementioned potential miRNAs should be investigated in more details. Mir-221-3p and miR-155-5p were induced in U2932-EBV cells, compared to U2932 cell line. These results were consistent with Lawrie et al. data [137]. Mir-221-3p was not detected in neither of SUDHL5-EBV or SUDHL5 cell line.

In the brain approximately 70% of the miRNA are expressed and most of it are neuron specific. MiRNA are important element for proper nervous system function. These miRNA plays a crucial role for brain function. Any disruption in the miRNA processing mechanism would cause diseases like ataxia in Purkinje cells, multiple sclerosis in oligodendrocyte cells, Parkinson’s disease in dopaminergic cells, Alzheimer’s disease in α-calcium/calmodulin-dependent protein kinase type II (αCaMKII)-expressing neurons (16).

The reason for this is that epigenetics can silence tumor suppressors all though that is negative side of epigenetics it is also positive, it can also turn off negative genes like cancer or family diseases known to be passed down. Tumor suppressors which protect abnormal cell growth can be turned off by epigenetics but with new research scientist are finding a way to silence certain epigenetics. In this new research scientist are finding ways to turn on or silence certain epigenetics, doing this could prevent someone from inheriting a family disease like factor 5 Leiden which in my case my mother has and could have passed on to me although I have been tested and I didn’t inherit that trait epigenetics could have silenced that trait off for me or it could be a recessive trait which means it would skip me but could be inherited by my children. That is why doctors are researching ways to silence or turn on certain traits, this could save many people from diseases known in your family or help someone turn on tumor suppressors that are there to help our cells from becoming

The eleven double-stranded RNA segments remain within the protection of the two protein shells. The RNA dependent RNA polymerase of the virus creates mRNA transcripts of the double-stranded viral genome. By remaining in the core, the viral RNA evades innate host immune responses known as RNA interference that are triggered by the presence of double-stranded RNA.

Science has led to the discovery of DNA and then identification of specific genes in organisms that control heredity and determine a cell 's function. With the advancement of research we are now able to explore the different roles of genes and understand how they might be correlated with diseases.

This mechanism inhibits translation of the protein for which the target mRNA codes for and therefore silences that specific gene. RNAi can also involve the use of coded short hairpin RNAs (shRNAs), which are larger dsRNA molecules containing a loop section which folds back on itself to form a double stranded molecule contains a sense and antisense strain. The shRNAs are cleaved into siRNAs when they bind to RISC and can also found in examples of exogenous introduction such as viral or experimental insertion. The ability to post translational control of gene expression with a precise sequence specific manner has been exploited experimentally in researching gene knockdown and loss of function gene analysis.

It has been reported in lung cancer tissues and cell line MIRL lncRNA acts as an oncogene. This downregulation is confirmed by knocking down MIR4435-2HG in lung cancer and treating the lung cancer cells with resveratrol. In both the cases,

1998). Interestingly, the phenomenon that the introduction of double-stranded RNAs “quelled” the expression of endogenous genes contain homologous sequences had been observed and even utilized as a tool to knock down gene expression by researchers long before the underlying RNAi mechanism was identified and separated from the antisense silencing by Fire and Mello in 1998 (Fire et al. 1998). They consistently found that the introduction of double-stranded RNAs (dsRNAs) was 100 or more folds effective than the single-stranded “sense” or “antisense” RNAs (ssRNA) to silence the complement gene expression (Fire et al. 1998; Kennerdell and Carthew 1998). Moreover, the dsRNA mediated gene silencing can be transferred between adjacent cells, and even inherited through multiple rounds of cell division (Fire et al. 1998). Therefore, it is proposed that this so called “RNA interference” is an inheritable and transferrable process that involves a catalytic process, which differs from the previously known antisense silencing that ssRNAs directly bind to the complement mRNA targets to repress translation by blocking ribosome access. Subsequently, Tuschl and co-workers demonstrated that RNAi mechanism exists in mammalian cell lines for the introduction of synthetic