The Role of MicroRNAs in Gene Regulation and Disease

 Introduction

In the ever-evolving field of biochemistry, one of the most exciting areas of research involves the study of microRNAs (miRNAs). These small, non-coding RNA molecules play a crucial role in regulating gene expression, impacting a wide range of biological processes and contributing to various diseases. In this blog post, we will explore the biogenesis of miRNAs, their function in gene regulation, and their implications in human health and disease.

Biogenesis of MicroRNAs

MicroRNAs are approximately 22 nucleotides long and originate from longer RNA precursors that are transcribed by RNA polymerase II. The biogenesis of miRNAs involves several steps:

1. Transcription: Primary miRNAs (pri-miRNAs) are transcribed from miRNA genes located in the genome.

2. Processing: The pri-miRNAs are cleaved by the microprocessor complex, consisting of the enzyme Drosha and its partner DGCR8, into precursor miRNAs (pre-miRNAs).

3. Exportation: Pre-miRNAs are exported from the nucleus to the cytoplasm by Exportin-5.

4. Dicing: In the cytoplasm, the enzyme Dicer further processes the pre-miRNAs into mature miRNA duplexes.

5. Incorporation: One strand of the miRNA duplex (the guide strand) is incorporated into the RNA-induced silencing complex (RISC), while the other strand (the passenger strand) is degraded.

Function in Gene Regulation

Once incorporated into the RISC, miRNAs guide the complex to target messenger RNAs (mRNAs) by base-pairing with complementary sequences, typically in the 3’ untranslated region (UTR) of the target mRNA. This binding can result in:

1. mRNA Degradation: The RISC can induce the degradation of the target mRNA, effectively reducing its levels in the cell.

2. Translational Repression: The RISC can inhibit the translation of the target mRNA without degrading it, leading to decreased protein synthesis.

3. Gene Silencing: Both mechanisms contribute to the overall effect of gene silencing, regulating the expression of genes at the post-transcriptional level.

Implications in Health and Disease

MicroRNAs are involved in numerous physiological processes, including development, differentiation, proliferation, and apoptosis. Dysregulation of miRNA expression has been linked to a variety of diseases, highlighting their importance in maintaining cellular homeostasis.

1. Cancer: Aberrant miRNA expression can lead to uncontrolled cell growth and cancer. For example, miR-21 is often upregulated in various cancers and acts as an oncogene by inhibiting tumor suppressor genes.

2. Cardiovascular Diseases: miRNAs such as miR-1 and miR-133 play critical roles in cardiac development and function. Altered levels of these miRNAs are associated with heart diseases, including myocardial infarction and heart failure.

3. Neurodegenerative Disorders: miRNAs are also implicated in neurodegenerative diseases. For instance, miR-29 and miR-9 are involved in Alzheimer’s disease by regulating the expression of proteins associated with amyloid plaque formation.

4. Immune Response: miRNAs like miR-155 are key regulators of the immune system, influencing the development and function of immune cells. Dysregulation of immune-related miRNAs can lead to autoimmune diseases and chronic inflammation.

Therapeutic Potential

Given their central role in gene regulation and disease, miRNAs are attractive targets for therapeutic intervention. Strategies to modulate miRNA activity include:

1. miRNA Mimics: Synthetic miRNAs that mimic the function of endogenous miRNAs can be used to restore the expression of downregulated miRNAs in diseases.

2. AntagomiRs: These are chemically modified anti-miRNA oligonucleotides designed to inhibit the function of specific miRNAs that are upregulated in diseases.

3. miRNA Sponges: These are RNA molecules with multiple miRNA binding sites that can sequester and inhibit specific miRNAs.

Conclusion

MicroRNAs are pivotal regulators of gene expression with significant implications in health and disease. Understanding the biogenesis, function, and regulatory mechanisms of miRNAs opens new avenues for diagnostic and therapeutic approaches in various medical conditions. As research in this field continues to advance, the potential for miRNA-based therapies to revolutionize medicine becomes increasingly promising.

WESTERN BLOT

A Western Blot (immunoblotting)is an analytical technique used for detection of specific proteins. The protein from the sample is first separated according to size by SDS-PAGE (Polyacrylamide Gel Electrophoresis) and then transferred to a nitrocellulose or Polyvinylidene Fluoride (PVDF) membrane where they become immobilized. Then they are detected first by a specific primary antibody and then by enzyme linked secondary antibody. The enzyme linked to the secondary antibody forms a colored band after conjugation which shows the presence of specific protein. It is used as a confirmatory test in many medical diagnostic tests like HIV, because it gives information if a specific antibody is been secreted against the pathogen.

Biochemistry

QUESTION: The Managing Director of a well-known company on Wall Street thrives on a diet of fruit jam, bread, pasta, and coffee. She exercises intermittently. One day she decides to go to her primary healthcare provider for a routine checkup. The healthcare provider recommends that she take the Benedict's test. Assume that the glucose levels of the patient are high. 1. State the results that the test would indicate (specify the color of the solution). 2. State the composition and the properties of the ketohexose derived from fruit jam. 3. Describe the manner in which ketohexose acts as a reducing sugar in the test.

ANSWER: 1. Benedict's test detects reducing sugars (such as glucose). Benedict's reagent contains blue copper ions (Cu2+) which are reduced to copper (Cu+) in the presence of reducing sugars. These are precipitated as red copper(I) oxide which is insoluble in water.Therefore the color of the solution would be Red.

2.Ketohexoses is a ketone containing hexose i.e. it consists of six carbon monosaccharides with a keto group ( R-CO_R ) and other carbon atoms consists an OH group and remaining hydrogens. Ketohexoses are soluble in , doesnt have sharp melting points, can char and sweetin taste because of which are added in the jam. 

3. When the hemi-acetal or ketal hydroxyl group is not engaged or locked or linked to another sugar molecule but  free then the aldehyde (or keto-) form (i.e. the chain-form) is available for reducing copper (II) ions. When a sugar is oxidized, its carbonyl group (i.e. aldehyde or ketone group) is converted to a carboxyl group.