Area Of Genetic Engineering Research Articles

Ethical Issues on Cloning and Genetically Modified Products

Science and Technology have greatly improved man’s quality of life as new discoveries are designed to nurture a healthy and long life. Science and Technology can be described as two sides of a coin as science discovers the problem and technology provides the product to solve that particular problem that affects the well-being of mankind. Philosophy, like science, is a discipline that is hinged on rational thinking and the pursuit of truth. The branch of philosophy that is concerned with how man ought to live in the society is Ethics. Ethics is the branch of philosophy that deals with rightness and wrongness of human actions. It also determines the rightness or wrongness of technological innovations and their effect on the health of man. There are numerous scientific discoveries in the area of genetic engineering but this paper is limited to cloning and genetic modification. In this work, ethical implications of these scientific discoveries were identified. This paper shall use the method of conceptual analysis. It intends to address the ethical implications of cloning and genetically modified products for mankind. The paper concludes that the benefits of these scientific developments should be embraced as they outweigh the cons and provide succour to beneficiaries which is the objective of Science and Technology. The welfare of mankind is the concern of ethics as well as science and technology.

A comprehensive overview of CRISPR/Cas 9 technology and application thereof in drug discovery.

Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-Cas technology possesses revolutionary potential to positively affect various domains of drug discovery. It has initiated a rise in the area of genetic engineering and its advantages range from classical science to translational medicine. These genome editing systems have given a new dimension to our capabilities to alter, detectand annotate specified gene sequences. Moreover, the ease, robustnessand adaptability of the CRISPR/Cas9 technology have led to its extensive utilization in research areas in such a short period of time. The applications include the development of model cell lines, understanding disease mechanisms, discovering disease targets, developing transgenic animals and plants, and transcriptional modulation. Further, the technology is rapidly growing; hence, an overlook of progressive success is crucial. This review presents the current status of the CRISPR-Cas technology in a tailor-made format from its discovery to several advancements for drug discovery alongwith future trends associated with possibilities and hurdles including ethical concerns.

Gene-independent therapeutic interventions to maintain and restore light sensitivity in degenerating photoreceptors.

Neurodegenerative retinal diseases are a prime cause of blindness in industrialized countries. In many cases, there are no therapeutic treatments, although they are essential to improve patients' quality of life. A set of disease-causing genes, which primarily affect photoreceptors, has already been identified and is of major interest for developing gene therapies. Nevertheless, depending on the nature and the state of the disease, gene-independent strategies are needed. Various strategies to halt disease progression or maintain function of the retina are under research. These therapeutic interventions include neuroprotection, direct reprogramming of affected photoreceptors, the application of non-coding RNAs, the generation of artificial photoreceptors by optogenetics and cell replacement strategies. During recent years, major breakthroughs have been made such as the first optogenetic application to a blind patient whose visual function partially recovered by targeting retinal ganglion cells. Also, RPE cell transplantation therapies are under clinical investigation and show great promise to improve visual function in blind patients. These cells are generated from human stem cells. Similar therapies for replacing photoreceptors are extensively tested in pre-clinical models. This marks just the start of promising new cures taking advantage of developments in the areas of genetic engineering, optogenetics, and stem-cell research. In this review, we present the recent therapeutic advances of gene-independent approaches that are currently under clinical evaluation. Our main focus is on photoreceptors as these sensory cells are highly vulnerable to degenerative diseases, and are crucial for light detection.

Application of Restriction Free (RF) Cloning in Circular Permutation.

The restriction free (RF) cloning has emerged as one of the highly efficient techniques in the area of genetic engineering. RF cloning has wide range of applications in plasmid DNA manipulation including cloning of a single gene, simultaneous assembly of multiple DNA fragments, and mutagenesis from single to multiple simultaneous alterations of a target DNA. Recently, we have developed a new technique of circular permutation using RF cloning. Circular permutation is widely used to investigate the mechanisms of protein folding and function. Previously, restriction enzyme based cloning was used to introduce circular permutation. Our RF cloning method made the protocol faster and more cost-effective. In this chapter, we describe a step-by-step protocol for generating circular permutants using RF methodology.

A Concise Review of Dendrocalamus asper and Related Bamboos: Germplasm Conservation, Propagation and Molecular Biology.

Bamboos represent an emerging forest resource of economic significance and provide an avenue for sustainable development of forest resources. The development of the commercial bamboo industry is founded upon efficient molecular and technical approaches for the selection and rapid multiplication of elite germplasm for its subsequent propagation via commercial agro-forestry business enterprises. This review will delve into the micropropagation of Dendrocalamus asper, one of the most widely cultivated commercial varieties of bamboo, and will encompass the selection of germplasm, establishment of explants in vitro and micropropagation techniques. The currently available information pertaining to molecular biology, DNA barcoding and breeding, has been included, and potential areas for future research in the area of genetic engineering and gene regulation have been highlighted. This information will be of relevance to both commercial breeders and molecular biologists who have an interest in establishing bamboo as a crop of the future.

CRISPR/Cas9 in epigenetics studies of health and disease.

Epigenetics is the heritable phenotypic changes without altering the genotype. Epigenetic processes are such as histone methylation, acetylation, ubiquitination, sumoylation, phosphorylation, ADP ribosylation, DNA methylation and non-coding RNAs interactions associated with structural changes in chromatin. The change of structure is either open chromatin for "active" state or closed chromatin for "inactive" state, that regulates important biological phenomenon like chromatin condensation, gene expression, DNA repair, cellular development, differentiation and homeostasis, etc. However, dysregulation of epigenetic patterns causes diseases like cancer, diabetes, neurological disorder, infectious diseases, autoimmunity etc. Besides, the most important clinical uses of Epigenetics studies are i. identification of disease biomarkers and ii. development of their therapeutics. Epigenetic therapies include epi-drugs, combinatorial therapy, nanocarriers, plant-derived products that are being used for changing the epigenetic pattern to reverse gene expression. However, the developed epi- drugs cause off-target gene and transposable elements activation; promote mutagenesis and carcinogenesis in normal cells, are the major hurdles regarding their clinical use. Therefore, advanced epigenetic therapeutics are required to develop target-specific epigenetic modifications to reverse gene expression pattern. CRISPR-Cas9 (Clustered Regularly Interspaced Palindrome Repeats-associated protein 9) system-mediated gene activation mechanism paves new methods of target-specific epigenetic therapeutics to cure diseases. In this chapter, we discuss how CRISPR/Cas9 and dCas9 have recently been engineered for epigenome editing. Different strategies have been discussed used for epigenome editing based on their efficacy and complexity. Last but not least we have discussed the limitations, different uses of CRISPR/Cas9 and dCas9 in the area of genetic engineering.

Methods of genome editing for increasing the shelf life of tomato fruit

Genome editing methods are now widely used in research aimed at studying fundamental biological processes, in particular for regulating maturation and extending shelf life of plant agricultural products. This review briefly discusses plant genome editing methods and examples of their successful application for increasing the storage life of fruits of tomato as one of the most important crops. Genome editing is one of the new areas of genetic engineering that is truly revolutionary in biotechnology. Various genome editing systems have been developed over the past decades: zinc finger nucleases (ZFNs), transcriptional activator-like effector nucleases (TALENs), and clustered regularly located short palindromic repeats recognized by Cas9 nuclease (CRISPR/Cas9). The most common and widely used is the CRISPR/ Cas9 system, which has many advantages over other existing genome editing systems.

Clinical practice: chimeric antigen receptor (CAR) T cells: a major breakthrough in the battle against cancer.

Chimeric antigen receptor (CAR) T cell therapy has come of age, offering a potentially curative option for patients who are refractory to standard anti-cancer treatments. The success of CAR T cell therapy in the setting of acute lymphoblastic leukemia and specific types of B cell lymphoma led to rapid regulatory approvals of CD19-directed CAR T cells, first in the United States and subsequently across the globe. Despite these major milestones in the field of immuno-oncology, growing experience with CAR T cells has also highlighted the major limitations of this strategy, namely challenges associated with manufacturing a bespoke patient-specific product, intrinsic immune cell defects leading to poor CAR T cell function as well as persistence, and/or tumor cell resistance resulting from loss or modulation of the targeted antigen. In addition, both on- and off-tumor immunotoxicities and the financial burden inherent in conventional cellular biomanufacturing often hamper the success of CAR T cell-based treatment approaches. Herein, we provide an overview of the opportunities and challenges related to thefirst form of gene transfer therapy to gain commercial approval in the United States. Ongoing advances in the areas of genetic engineering, precision genome editing, toxicity mitigation methods and cell manufacturing will improve the efficacy and safety of CAR T cells for hematologic malignancies and expand the use of this novel class of therapeutics to reach solid tumors.

Molecular Dynamics Studies of Human β-Glucuronidase

Problem statement: The enzyme β-glucuronidase is being used as a reporter molecule in the area of genetic engineering, as a component of prodrug therapy in cancer treatment and in the scouring process of cotton fabrics. However, a detailed understanding of the factors responsible for the stability and the activity of this enzyme is still not available. Molecular Dynamics (MD) simulations provide an estimate of equilibrium and dynamic properties of enzyme systems that cannot be calculated analytically. With this perspective, molecular dynamics simulations of human β-glucuronidase (GUS) have been carried out to determine the behavior of this enzyme in vacuum and solvent environments at a defined temperature. Approach: CHARMM force field along with distance dependent dielectric model was used to represent the solvent environment in the MD simulations. The parameters employed in various stages of MD simulations had been selected based on repeated trials under various conditions as a method of choosing the optimum parameters for each stage. Results: It was found that simulations in vacuum caused the backbone of GUS to have smaller fluctuations from their mean values compared with the fluctuation in implicit solvent simulations, due to the fact that vacuum environment did not provide for the electrostatic interactions affecting the backbone of GUS that may otherwise exist in a solvent environment. Conclusion: Inclusion of solvent effects in MD simulations is crucial in understanding structural flexibility and stability of β-glucuronidase. Implicit solvent method can provide a realistic inclusion of backbone flexibility and structural compactness of GUS, which will have profound influence on the stability and activity of the enzymes, with a marginal increase in computational time.

Optimization of in vitro micropropagation and regeneration for Populus × interamericana and Populus × euramericana hybrids (P. deltoides, P. trichocarpa, and P. nigra)

A rapid and efficient micropropagation method has been established for six European poplar cultivars of economic interest – four Populus × interamericana and two Populus × euramericana. Using a three-step procedure, we were able to regenerate plantlets from callus and acclimate them within 4 months. In the first step, callogenesis was induced when explants were cultured for 25 days on culture medium supplemented with 10 µM α-naphthaleneacetic acid and 5 µM N6(2-isopentenyl)adenine. Bud regeneration followed by shoot elongation was then obtained from callus tissue by combining the cytokinin-like compound thidiazuron with the surfactant Pluronic F-68 at concentrations adjusted for each cultivar. The usefulness of this procedure in the area of genetic engineering is discussed.

Rereading Christa Wolf's "Selbstversuch": Cyborgs and Feminist Critiques of Scientific Discourse

In her seminal Cyborg Manifesto, Donna Haraway invokes the cyborg, that hybrid of organism and machine, as a figure for a subversive feminist practice. Haraway breaks with traditional feminist approaches that have relied on Western origin myths and dualistic thought.1 Instead of rejecting or avoiding the male-dominated fields of technology and science, Haraway, by embracing the figure of the cyborg, calls on women to actively shape scientific and technological developments. This call resonates with Christa Wolf's story Selbstversuch, in which a female scientist participates in and critically comments on a biochemically induced sex-- change experiment. Wolf's story combines elements of science fiction and bio-technological discourse-two areas in which cyborgs abound. Thus Selbstversuch provides an ideal framework for critical reflections on the relationship between gender and science in general and on Haraway's feminist reading of the cyborg figure in particular.2 The main protagonist in Selbstversuch assumes a double role: she is a scientist who prepares and subsequently undergoes the experiment, and who writes the classified research report during the experiment (this official report is merely alluded to in the text); and she is the first-person narrator and of the story Selbstversuch: zu einem Protokoll-a on the official scientific report.3 She writes this treatise in hindsight, i.e., after breaking off the experiment and transforming herself back into a woman. In this critical commentary, ironic and personal in tone, she addresses her professor, who functions rhetorically also as the embodiment of male-dominated scientific discourse. The reference in the story's subtitle to a Traktat (treatise)-traditionally representing a scientific or philosophical argument-illustrates the narrator's objective: to challenge established scientific conventions from within. The narrator's ironic comment, [es gibt] nichts Komischeres [...] als Frauen, die Traktate schreiben (SV 86-87;4 there is nothing funnier than women who write treatises, SE 215) indicates the social sanctions (ridicule) for female authorship of a male-dominated genre and hence the transgressive character of her undertaking. Supplementing the classified research report with a treatise penned by a female author accomplishes three things. First, by adopting the perspective of the usually silenced subject of the experiment, this appendix questions the exclusive truth claim of the scientist-embodied in the official report. Second, the treatise gives voice to another traditionally silenced subject position: that of the female scientist who no longer adheres to an androcentric position. Thus the female subject/scientist takes control not only of her body (by breaking off the experiment), but also of the way the experiment is interpreted. Third, by breaking the secrecy code of the classified report, the treatise brings out into the open the moral void left by the report. As I will illustrate later in the article, the treatise ultimately calls for a different approach to the sciences, one that considers the ethical and social implications of bio-technological research not as separate from but as part and parcel of its epistemological foundation. Wolf wrote Selbstversuch in 1972(5), situating the story twenty years into the future-in the year 1992. Yet from today's perspective, the story takes place in the recent past and the experiment has indeed lost some of its science-fiction dimensions due to advances in the field of bio-technology, especially in the areas of genetic engineering and reproductive technologies. Even the 1989 film adaptation of Selbstversuch produced by DEFA and directed by Jochen Vogel in the last year of the GDR's existence demonstrates the extent to which reality has caught up with the sci-fi character of the original text. The only part of the film that remains unreal is the technology of the actual sex change (induced through a series of injections). …

Human Gene Therapy: Ethics and Public Policy

The first three human gene transfer/therapy clinical protocols are now underway after having been subjected to an extensive review process by the Recombinant DNA Advisory Committee (RAC) and its Human Gene Therapy Subcommittee. The "Points to Consider" document developed by the RAC established the framework for evaluating genetic intervention protocols. This review process is taking place in a broader social context. Public attitude surveys in this country have indicated a general lack of knowledge in the area of genetic engineering but an acceptance of somatic-cell gene therapy as treatment for disease. Internationally, numerous policy statements on human genetic intervention have been published, all of which support the moral legitimacy of somatic-cell gene therapy for the cure of disease. The debate over the ethical issues related to somatic-cell gene therapy has evolved over a ten-year-period. The time has now come to begin a formal public process for the ethical assessment of germ-line genetic intervention.

Reflections on the Limits of Science and Technology

SYNOPSIS. Exuberance over insights gained in the infant field of genetics early this centuryled scientists to extrapolate beyond their data to heredity of behavioral traits in people. Oneof the direct consequences was the incarceration of Americans and Canadians of Japanese ancestry during World War II as enemy aliens. Drawing on this personal experience of the misapplication of science, I describe the process of scientific indoctrination and blindness to the limitations of this way of knowing. This led to my attempt to demystify science through the electronic media. Only recently have I come to understand that two assumptions that impelled me to use television in the first place are wrong. The first was that with access to more information about science, the general public would be in a position of making better informed decisions on issues involving science and technology. The problem is that we are overwhelmed with information and most people lack the ability to distinguish meaningful “signal” ( i.e. , credible information) from background “noise” ( i.e. , garbage). We believe in phantoms created by the acceptance of anything because it exists as print or television programs. My second assumption had been that we need a mechanism to do an in-depth “cost/benefit” analysis of new technologies before they are actually made available. But history reveals that the benefits of new technologies are immediate and obvious while the costs are usually hidden and completely unpredictable. But in the rush to exploit new scientific insights we ignore the fact that science must lookat nature in isolated bits and pieces. Knowledge gained in fragments does not yield an understanding of the greater context from which the pieces are taken. With each new discovery, we itch to apply it, forgetting how much we have yet to learn. Our attempts to manipulate nature are often illusions of control created by our ability to overpower nature by brute strength. In the area of genetic engineering, this could be truly disastrous.

Roundtable discussion of research priorities.

The purpose of this panel is to discuss some future research needs and priorities in the area of genetic engineering of microorganisms.