MICR40010 Recombinant DNA Technology

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MICR40010 Recombinant DNA Technology

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MICR40010 Recombinant DNA Technology

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Course Code: MICR40010
University: Trinity College Dublin

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Country: Ireland


Topic: Bio-Therapeutic For Drug Addiction
Should Only Use R DNA Technology to Solve the Problem
A Biotechnology company has become interested in developing a commercially viable immunotherapy to help treat drug addiction. You have been asked to investigate this possibility and present the approach which would be most likely to succeed. 


Introduction to Therapeutic and Drug Addiction
Since 20th-century Drugs have been the biggest issue in society.  Drugs by definition are the chemical substance that is used in treatment. It is something that has an impact on everyone, despite race or financial status. They affect both how society is shaped and the users. Precisely there are hundreds of drugs presently available. All apparently have a similar effect on the user despite potent or illegal; they negatively affect the body and the mind. Other than these bad effects, there is the financial condition that has afflicted users over many generations. Rich or poor, chemicals are required by the body and the user for the feel of extreme high. Beyond all, there is a criminal aspect that mostly comes in play in the society. According to the studies carried out over the past decades gives evidence which gravitates to show that drug is the main factor for some people to commit illicit acts.
This primarily leads to another drug usage, mainly harder drugs like heroin and benzodiazepines which carries greater effects on the human body. According to Banducci, Hoffman and et al hard drugs lead to the withdrawal of human emotions like dysphoria, physical pain, alexithymia, loss of motivation and malaise (Banducci, Hoffman, Lejuez, & Koenen, 2014). Regarding Banducci and et al, these emotions are associated with molecular neuro-adaptive changes inside the nucleus accumbens. As a result, it decreases the functioning of the mesocorticolimbic dopamine system.
Nestler explains how continuous drug use can lead to addiction and how it changes the brain. According to Nestler, the brain is changed by continuous drug exposure, even if is self-administered or not (Nestler, 2014). Statistically, it is obvious that self-administration of addictive drugs causes the peripheral alterations in the brain beyond those achieved by passive receiving of the drug. In the case of smokers, they undergo primary stimulus for yearning (Wise & Koob, 2014).
Regular use of nicotine and alcohol according to the research by Nestler alters the gene expressions. And as a result, it leads to genetic variations in serotonin transporter that enhances psychiatric disorder (EJ Nestler, 2014). Through Nestler research concludes that drug and drug addictions have many negative impacts on society. But these problems can be solved by the use of R-DNA Technology.
Introduction to R DNA Technology
In the last decades, R DNA technology was believed by scientist that the end result of the technology would result in undesirable characteristics by the experimented organism. However, in the recent era, it has shown promising result and, in fact, it is highly credited for the advancement of modern medicine. Additionally, due to its advancement in the medical sector, it has led to cloning technology, Transcription Technology, gene knock out technology and GENETECH. However, only Gene Therapy and Gene Knock out will be discussed in this paper and how technology can curb or reduce drug addiction.
Gene Therapy 
Gene Therapy Is a progressive technique with therapeutic potential in the health sector. The first successful report on the gene therapy was when it provided the safest way of treating genetic disease. The strategy showed a good response to the treatment of adenosine deaminase-deficiency (ADA-SCID). At the begging, the technology encountered several challenges like the upkeep of patients on PEGylated ADA (PEG-ADA) during gene therapy. Reason for the unsuccessful result was due to the targeting of gene transfer to T-lymphocytes.  However, successful results were obtained hematopoietic stem cell (HSCs) was targeted.
Disorders like adrenoleukodystrophy (X-ALD) and X- linked are likely through the expression of explicit genes transferred by a lentiviral vector. X-ALD protein expression showed shows the successful achievement of gene-correction of true HSCs. The use of a lentiviral vector was successfully made to treat genetic human disease for the first time. In 2006, metastatic was treated through immunotherapy through the enhancement of specific protein expressions. This success led to the treatment of serious death causing disease through immunotherapy in the field of health sciences (Merten & Gaillet, 2016).
The transfer of genes to a smaller number of cells at anatomically separates sites in a targeted strategy that has the capability to confer the therapeutic advantage. Impressive results were shown for incurable autosomal recessive dystrophies. In April 2006, Swiss-German gene therapy clinical trial aimed to treat chronic granulomatous came up with success (Godinho, Aguinaga, Grynszpan, Lima, Azulay, Cuzzi, & Manela?Azulay, 2015)
Gene therapy has also targeted many different cancers including, neurological, skin gynecological as well as pediatric tumors and hematological malignancies. Different strategies like inserting a tumor suppressor gene to immunotherapy, oncolytic virotherapy, and prodrug therapy have been used to treat cancer. The key player in the cancer treatment is the p53 gene transfer. In some approaches, p53 gene transfer is combined with radiotherapy or chemotherapy. The most important strategies that have used until now are vaccination with tumor cells (Scott, Bayly, Abell, & Skidmore, 2016). Vectors like new fiber chimeric oncolytic adenovirus (Ad5/35-EGFP) were transduction improvement as more progeny virus was created in HCC. This technology inhibited the growth of a tumor. Currently, cancer therapy has become effective and more advanced compared to recent years.
Another gene therapy strategy in health science is the treatment of cardiovascular disease. Gene therapy will give a new venue for myocardial protection, therapeutic angiogenesis prevention of bypass graft failure, risk factor management, and prevention of restenosis following angioplasty. Wiskott – Aldrich syndrome which is an inherited immunodeficiency is caused by the regulating of protein in the cytoskeleton. Its treatment needs stem cells transplantation and if the donors are unavailable gene therapy is carried out through an infusion of modified HSPC ex vivo (Hernandez, Padilla, Dakhel, Coll, Hervas, Adan, Masa, Mitjans, Martinez, Coma &Rodríguez, 2013).
Genes knock out Technology 
In essence, Genes knock out Technology is another progressive technique with therapeutic potential in the health sector.  A gene knockout is a technique where organisms are engineered the imperative genes that have been made. This technique is accomplished by combining several techniques through various procedures. The ultimate objective of gene knock out technology is to disclose the function of a gene discovered by large-scale sequencing projects. For instance, in 1996 the international collaboration of scientists from United States, Europe, and Japan started genome sequencing of Columbia strain (Col-0) which was completed in 2000. Through gene knock out technique the first world blue rose was created by Melbourne Biotechnology Company licensed Florigene Ltd in 2008.  The Japanese and Australia researchers have explained the application of RNAi technology for gene replacement in plants, creating the world’s only blue rose.
In gene knock out technology has been several cases in alcohol research, mutation isolation and lastly on how to reduce carcinogens in tobacco leaves. In this case, only cases of alcohol research and how to reduce carcinogens in tobacco will be considered. Alcoholism is a complex condition that involves various disorder and behavioral characteristics.  Human and animals studies showed how the aspects of alcoholism are mostly mediated Neurotransmitter system (Neuschwander & Loomba, 2015). Neuroscientists’ main goal is to determine gene and protein in the brain that impacts the expression of alcoholism. The enhancement of inhibition and initiation of reward pathways are among the activity of the brain that is affected by alcoholism.
The in other case, scientists have discovered how carcinogen in tobacco leaves can be reduced by suppressing specific gene. According to them, it may result in a tobacco product with a minimal amount of cancer-causing agent, especially to smokeless tobacco products. The demethylase gene is deactivated which changes nicotine into non-nicotine as the tobacco is cured, processed and stored.
In the production of recombinant pharmaceutical microbial cells are used mostly which shows that many obstacles come into the way by restricting the efficiency of the production of functional proteins. The efficient production of functional proteins is controlled with alterations in the cellular system. Cell stress activation, posttranslational modifications, low solubility, and resistance in expressing new genes are the obstacles that must be dealt with. Mutation taking place in human at genetic level sources deficiencies in production of protein, which can be treated by the combination of external genes to fill the gap and reach the required level.
Recombinant DNA research indicates huge promise in the further apprehension of yeast Biology by creating possible the manipulation and analysis of yeast genes. The advancements in technology have made the analysis and genetic manipulation in the yeast to be feasible (Wu, Zhao, Ma Mitchell, Xie, Lin, Wang, Xiao, Wang, Zhoua, &Liu, 2017). The R DNA technology is currently passing rigorous development which has led to great changes in the research lines. This rigorous development has also opened management of research for biosynthetic pathways. In pharmaceutical production, actinomycetes are used. For example, the manipulation of biosynthetic pathways and useful compounds in health science advance drug generation.
Implementation of phage Display Technology
Due to the fact that genotype phage and phenotype phage are linked directly, the technology can be used to develop a vaccination to people with drug addiction. Actually, the resulting link between the phenotype and the genotype is a result of the insertion of a gene encoding, a protein of importance to a phage protein gene. For this reason, when a bacterial host is infected by phage particles and conveys their genes, the encoded guest amino acid by unknown DNA is indicated as a part of the admissible coat protein.  As a result, the surface of phage virion exposes the displayed fusion protein.
The functional peptides with the desired features are selected in an effective and straightforward manner. However, for the construction of a phage display library, randomized oligonucleotide sequences are joined on a phage protein gene.  Since the entire display library has a large number of peptides, the phage particles chosen from the library with its displayed peptides will be propagated into a host cell as virion acts as independent vector and is able of infecting the bacteria. Accordingly, a large quantity of peptides of viral particles is screened to get the most favorable binders with the highest affinity to the target.
Instead of engineering different peptides at one time at a time and analyzing each variant, phage display library will construct the required variant since it contains up to 1010 variants. The ability of phage particles to outstand low PH and low temperatures had allowed the foundation of using low pH and higher concentration urea to be used to dissociate bound phage from a target.
The bacteriophage like Fd phage particles that contain a capsid which is a 930nm and 6.5nm diameter, composed of 11 genes enclosing a single-stranded DNA genome nucleotides. In addition, M13, F1, and Ft are the other examples of bacteriophage. Furthermore, the viral mass is estimated to be 16.3 MDa, composes mainly of 2700 copies of pVIII, a 50 a residue encoded by the gene. However, there are 5 copies of pIII and 2 copies of pVI. Moreover, pIII, 406 consists of an adsorption protein used to peptide the expression. Primarily, pIII has two functional domains.  The first function is to integrate the parts of the cuspid structure by using C-terminal while the second function is to bind the f pilus through an exposed N-terminal domain.
The E.coli which is a Gram-negative bacteria will be infected by fd particle that will lead to filamentous phage infection which will induce a stage in which a bacteria generate particles into a growing media. The infection will start with the addition of phage pIII to the F pilus of male E. coli.  The circular stranded DNA will enter into the bacteria where it would be changed by the host DNA replication machinery into a double-stranded plasmid replicate state.  Through rolling circle replication, the protein PIII and PVIII are formed by rolling circle process by which replicative form makes a single-stranded DNA.
The choice of a living cell will be done by either cell suspension or by adherent cell where unbound phage must be washed away and recovery is done through bacterial infection.  The peptides designated into bio-panning will be tested according to their capacity to normal and malignant urothelial cells.
In vivo selection will be used for depletion of peptide recognizing unspecified molecules from circulation and to identify the receptors selectively on cells. These receptors will molecular target development techniques and target therapy on cocaine. In regard to selective infective phage (SIP), the lacking N-terminal domain necessary for phage infectivity will be adapter molecules to provide a terminal of pIII to the phage.  The phage particles with 2 to 4 amino acid and pIII coal protein will be replaced by landscape phage. Therefore, clones with nanomolar affinity will be chosen from the phage library against their immobilized target.
The fusion protein generated by rules of liking the phage phenotype. Therefore, cMDA library will be displayed on the phage surface.  The phage clones that stick to IgE will be chosen by screening and enrichment of phage libraries. The enhancement of gene therapy vector by picking alternative ligands will raise the selectivity and, therefore, reduce the toxicity of gene therapy. As a result, the combination of gene delivery will take place. Henceforth, the joining of gene delivery technique and power of combinatorial phage libraries will allow the creation of genetically changed phage, displaying a known gene that will target ligand.  Through the result which is efficient and safer gene transfer into mammalian cells could lead to the desired gene that could lead to prevention of drug misuse that leads to addiction.
R DNA technology is a significant growth in science as it made the human life to be much easier. In recent years, it has furthered strategies for a biomedical application like cancer treatment, genetic diseases, and several plants disorders mostly in fungal and viral resistance. The role of R DNA technology is enhancing a clean environment and increased the resistance of a plant to various unfavorable acting factors. The developments it brought not only to plants and microorganisms but also to human is very important. The problems in enhancing the product at gene level occasionally face serious complications which are required to be dealt with for the benefit of R DNA technology future. In pharmaceutical, particularly, there are changes brought to genes are refused by the body hence hindering the production of good quality. Considering health issues, the R DNA technology is aiding in treaties some diseases which cannot be cured in normal circumstances.
Recombinant DNA technology has helped in reduction of drugs addiction. Some plants like tobacco have been genetically modified to reduce the negative effect on the human body. These advancements highly reduced the infection of diseases that are as a result of drug addictions like lung cancer and cardiovascular diseases. Scientists have been working to enhance vaccines and the production of the new ones by R DNA technology. These DNA vaccines which use the R DNA technology may be the key in highly reducing the use of drugs which are misused by several addictions. This modern vaccine initiates a small piece of disease into the body so that the body can create ways to fight that exact disease. It is evident that plants which are genetically engineered can cross breed with a wild plant, hence spreading their genes into the environment, polluting our biodiversity. Further worries exit that genetic engineering has hazard health effects. Thus, more extensive research is needed in this field to solve such problems and rectify the worries so as to make the world a better place for all common people.
Banducci, A. N., Hoffman, E. M., Lejuez, C. W., & Koenen, K. C. (2014). The impact of childhood abuse on inpatient substance users: Specific links with risky sex, aggression, and emotion dysregulation. Child abuse & neglect, 38(5), 928-938.
Godinho, M. M., Aguinaga, F., Grynszpan, R., Lima, V. M., Azulay, D. R., Cuzzi, T. & Manela?Azulay, M. (2015). Granulomatous reaction to red tattoo pigment treated with allopurinol. Journal of cosmetic dermatology, 14(3), 241-245.
Hernandez, J.L., Padilla, L., Dakhel, S., Coll, T., Hervas, R., Adan, J., Masa, M., Mitjans, F., Martinez, J.M., Coma, S. and Rodríguez, L. (2013). Therapeutic targeting of tumor growth and angiogenesis with a novel anti-S100A4 monoclonal antibody. PloS one, 8(9), e72480.
Merten, O. W., & Gaillet, B. (2016). Viral vectors for gene therapy and gene modification approaches. Biochemical engineering journal, 108, 98-115.
Nestler, E. J. (2013). Cellular basis of memory for addiction. Dialogues in clinical neuroscience, 15(4), 431.
Nestler, E. J. (2014). Epigenetic mechanisms of drug addiction. Neuropharmacology, 76, 259-268.
Neuschwander-Tetri, B. A., Loomba, R., Sanyal, A. J., Lavine, J. E., Van Natta, M. L., Abdelmalek, M. F., … & Kowdley, K. V. (2015). Farnesoid X nuclear receptor ligand obeticholic acid for non-cirrhotic, non-alcoholic steatohepatitis (FLINT): a multicentre, randomized, placebo-controlled trial. The Lancet, 385(9972), 956-965.
Scott, D. E., Bayly, A. R., Abell, C., & Skidmore, J. (2016). Small molecules, big targets: drug discovery faces the protein-protein interaction challenge. Nature Reviews Drug Discovery, 15(8), 533.
Wise, R. A., & Koob, G. F. (2014). The development and maintenance of drug addiction. Neuropsychopharmacology, 39(2), 254.
Wu, Y., Li, B.Z., Zhao, M., Mitchell, L.A., Xie, Z.X., Lin, Q.H., Wang, X., Xiao, W.H., Wang, Y., Zhou, X. and Liu, H. (2017). Bug mapping and fitness testing of chemically synthesized chromosome X. Science, 355(6329), eaaf4706.

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