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Description
A coincidence reporter construct, consisting of the p21-promoter and two luciferase genes (Firefly and Renilla), was constructed for the screening of drugs that might inhibit Olig2's tumorigenic role in glioblastoma. The reporter construct was tested using an Olig2 inhibitor, HSP990, as well as short hairpin RNA targeting Olig2. Further confirmatory

A coincidence reporter construct, consisting of the p21-promoter and two luciferase genes (Firefly and Renilla), was constructed for the screening of drugs that might inhibit Olig2's tumorigenic role in glioblastoma. The reporter construct was tested using an Olig2 inhibitor, HSP990, as well as short hairpin RNA targeting Olig2. Further confirmatory analysis is needed before the reporter cell line is ready for high-throughput screening at the NIH and lead compound selection.
ContributorsCusimano, Joseph Michael (Author) / LaBaer, Joshua (Thesis director) / Mangone, Marco (Committee member) / Mehta, Shwetal (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2014-05
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Description
Virus-Like Particles (VLPs) are self-assembling structures that lack the viral genetic material. Therefore they are safer and more immunogenic than other forms of vaccines. The Hepatitis B core (HBc) VLPs are a novel mechanism through which delivery of DNA-based human vaccines are plausible. Production of VLPs require recombinant, rapidly replicating,

Virus-Like Particles (VLPs) are self-assembling structures that lack the viral genetic material. Therefore they are safer and more immunogenic than other forms of vaccines. The Hepatitis B core (HBc) VLPs are a novel mechanism through which delivery of DNA-based human vaccines are plausible. Production of VLPs require recombinant, rapidly replicating, plant-based systems such as the geminiviral replicon system. This project entails the cloning process of HBc-DIII fusion protein, a VLP that should form Domain III of the Envelope protein on West Nile Virus, into deconstructed geminiviral vector. The cloning process includes the HBc-DIII fusion protein DNA isolation, restriction enzyme digestion with NcoI and SacI, PCR changing the NcoI site on the HBc-DIII insert to XbaI, sequencing, ligation into geminiviral vector and transformation into an agrobacterium strain. The major impediment to the cloning process was the presence of multiple bands instead of the expected two bands while doing restriction enzyme digests. The troubleshooting process enabled speculating that due to the excess of restriction enzymes in the digestion volume, some of the DNA was not digested completely. Hence, multiple bands were observed. However, sequencing analysis and further cloning process ensured the presence of HBc-DIII insert band (approximately 800bp) in the Gemini vector. Lastly, the construct HBc-DIII in Gemini vector was ensured to be in agrobacterium for further experiments such as agro-infiltration.
ContributorsSuresh Kumar, Reshma (Author) / Chen, Qiang (Thesis director) / Zhang, Peiming (Committee member) / School of Molecular Sciences (Contributor) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
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Description
In 2004, the South Korean geneticist Woo-Suk Hwang published what was widely regarded as the most important research result in biotechnology of the year. In the prestigious American journal Science, he claimed that he had succeeded in cloning a human blastocyst, an embryo in its early stages (Hwang et al.

In 2004, the South Korean geneticist Woo-Suk Hwang published what was widely regarded as the most important research result in biotechnology of the year. In the prestigious American journal Science, he claimed that he had succeeded in cloning a human blastocyst, an embryo in its early stages (Hwang et al. 2004). A year later, in a second Science article, he made the earth-shattering announcement that he had derived eleven embryonic stem cell lines using his cloning technique (Hwang et al. 2005). The international scientific community was stunned. American scientists publicly fretted that President George W. Bush‘s 2001 executive order limiting federal funding for stem-cell research in the United States had put American bioscience behind the Koreans‘ (Paarlberg 2005). These breakthroughs offered potential solutions to immune system rejection of transplanted organs and possible cures for diseases such as rheumatoid arthritis, Parkinson‘s, Down‘s syndrome, and paralysis (Svenaeus 2007). However, within a year, Hwang was exposed as a fraud who had faked his results and pressured his female colleagues to donate eggs without informed consent. Despite protests against his methods from Korean religious and nongovernmental organizations, Hwang had used his prestige to ignore his ethical obligations. The Korean government, too, was slow to investigate Hwang and to subject his work to appropriate regulation.
ContributorsClay, Anne (Author) / Hurlbut, James (Thesis director) / Maienschein, Jane (Committee member) / Marchant, Gary (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor)
Created2012-12
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Description
Reproductive cloning is the duplication of genetic material to reproduce a living organism. The sheep Dolly was the first adult mammal to be cloned and her birth unveiled a multitude of questions about the potential for cloning humans and how that might threaten human individuality. Given those questions, my project

Reproductive cloning is the duplication of genetic material to reproduce a living organism. The sheep Dolly was the first adult mammal to be cloned and her birth unveiled a multitude of questions about the potential for cloning humans and how that might threaten human individuality. Given those questions, my project delves into how reproductive cloning relates to the idea of individuality across three subgroups: humans, utility animals such as those used for research or agriculture, and pets.
ContributorsO'Connell, Lindsey Marie (Author) / Maienschein, Jane (Thesis director) / Ellison, Karin (Committee member) / Hurlbut, Ben (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor)
Created2013-05
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Description

Extrachromosomal circular DNA (eccDNA) has been identified in a broad range of eukaryotes and have been shown to carry genes and regulatory sequences. Additionally, they can amplify within a cell by autonomous replication or reintegration into the genome, effectively influencing copy number in cells. This has significant implications for cancer,

Extrachromosomal circular DNA (eccDNA) has been identified in a broad range of eukaryotes and have been shown to carry genes and regulatory sequences. Additionally, they can amplify within a cell by autonomous replication or reintegration into the genome, effectively influencing copy number in cells. This has significant implications for cancer, where oncogenes are frequently amplified on eccDNA. However, little is known about the exact molecular mechanisms governing eccDNA functionality. To this end, we constructed a fluorescent reporter at an eccDNA-prone locus of the yeast genome, CUP1. It is our hope that this reporter will contribute to a better understanding of eccDNA formation and amplification within a cell.

ContributorsKeal, Tula Ann (Author) / Wang, Xiao (Thesis director) / Tian, Xiaojun (Committee member) / School of Life Sciences (Contributor, Contributor) / Barrett, The Honors College (Contributor)
Created2021-05
Description

Polyketides are a wide ranging class of natural microbial products highly relevant to the pharmacological industry. As chemical synthesis of polyketides is quite challenging, significant effort has been made to understand the polyketide synthases (PKSs) responsible for their natural production. Native to Streptomyces, the aln biosynthetic gene cluster was recently

Polyketides are a wide ranging class of natural microbial products highly relevant to the pharmacological industry. As chemical synthesis of polyketides is quite challenging, significant effort has been made to understand the polyketide synthases (PKSs) responsible for their natural production. Native to Streptomyces, the aln biosynthetic gene cluster was recently characterized and encodes for an iterative type I polyketide synthase (iT1PKS). This iT1PKS produces both , and ,-double bond polyketides named allenomycins; however, the basis in which one bond is chosen over the other is not yet clear. The dehydratase domain, AlnB_DH, is thought to be solely responsible for catalyzing double bond formation. Elucidation of enzyme programming is the first step towards reprogramming AlnB_DH to produce novel industrially relevant products. The Nannenga lab has worked as collaborators to the Zhao lab at the University of Illinois at Urbana-Champaign to unravel AlnB_DH’s structure and mechanism. Here, mutant constructs of AlnB_DH are developed to elucidate enzyme structure and provide insight into active site machinery. The primary focus of this work is on the development of the mutant constructs themselves rather than the methods used for structural or mechanistic determination. Truncated constructs were successfully developed for crystallization and upon x-ray diffraction, a 2.45 Å resolution structure was determined. Point-mutated constructs were then developed based on structural insights, which identified H49, P58, and H62 as critical residues in active site machinery.

ContributorsBlackson, Wyatt (Author) / Nannenga, Brent (Thesis director) / Nielsen, David (Committee member) / Barrett, The Honors College (Contributor) / Chemical Engineering Program (Contributor) / School of Molecular Sciences (Contributor)
Created2023-05
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Description

Karl Oskar Illmensee studied the cloning and reproduction of fruit flies, mice, and humans in the US and Europe during the twentieth and twenty-first centuries. Illmensee used nuclear transfer techniques (cloning) to create early mouse embryos from adult mouse cells, a technique biologists used in later decades to help explain

Karl Oskar Illmensee studied the cloning and reproduction of fruit flies, mice, and humans in the US and Europe during the twentieth and twenty-first centuries. Illmensee used nuclear transfer techniques (cloning) to create early mouse embryos from adult mouse cells, a technique biologists used in later decades to help explain how embryonic cells function during development. In the early 1980s, Illmensee faced accusations of fraud when others were unable to replicate the results of his experiments with cloned mouse embryos. Illmensee also worked with human embryos, investigating how embryos split to form identical twins.

Created2017-02-26
Description

In the second half of the
twentieth century, scientists learned how to clone organisms in some
species of mammals. Scientists have applied somatic cell nuclear transfer to clone human and
mammalian embryos as a means to produce stem cells for laboratory
and medical use. Somatic cell

In the second half of the
twentieth century, scientists learned how to clone organisms in some
species of mammals. Scientists have applied somatic cell nuclear transfer to clone human and
mammalian embryos as a means to produce stem cells for laboratory
and medical use. Somatic cell nuclear transfer (SCNT) is a technology applied in cloning, stem cell
research and regenerative medicine. Somatic cells are cells that
have gone through the differentiation process and are not germ
cells. Somatic cells donate their nuclei, which scientists
transplant into eggs after removing their nucleuses (enucleated eggs).
Therefore, in SCNT, scientists replace the nucleus in an egg cell
with the nucleus from a somatic cell.

Created2014-11-04
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Description

The Boys from Brazil is a science fiction film based on the novel of the same name by Ira Levin about an underground neo-Nazi society in South America trying to clone Adolf Hitler, the dictator of Nazi Germany during World War II, to restore the Nazi movement. The film was

The Boys from Brazil is a science fiction film based on the novel of the same name by Ira Levin about an underground neo-Nazi society in South America trying to clone Adolf Hitler, the dictator of Nazi Germany during World War II, to restore the Nazi movement. The film was directed by Franklin Schaffner and released in 1978 by 20th Century Fox in Los Angeles, California. The Boys from Brazil is a film that was one of the first films to depict cloning, and to discuss the ethical implications of genetic engineering, cloning, and eugenics.

Created2017-06-09
Description

Advanced Cell Technology (ACT), a stem cell biotechnology company in Worcester, Massachusetts, showed the potential for cloning to contribute to conservation efforts. In 2000 ACT researchers in the United States cloned a gaur (Bos gaurus), an Asian ox with a then declining wild population. The researchers used cryopreserved gaur skin

Advanced Cell Technology (ACT), a stem cell biotechnology company in Worcester, Massachusetts, showed the potential for cloning to contribute to conservation efforts. In 2000 ACT researchers in the United States cloned a gaur (Bos gaurus), an Asian ox with a then declining wild population. The researchers used cryopreserved gaur skin cells combined with an embryo of a domestic cow (Bos taurus). A domestic cow also served as the surrogate for the developing gaur clone. The successful procedure opened the opportunity to clone individuals from species for which there are few or zero live specimens. The official release of this experiment's data was published in the paper 'Cloning of an Endangered Species (Bos gaurus) Using Interspecies Nuclear Transfer,' in October 2000. In the article, the researchers presented data collected from several cloned fetuses that were aborted before the full term of 283 days. At the time of publication, the gaur bull fetus, named Noah at birth, had developed for greater than 180 days. Noah was born on 8 January 2001, but died two days later due to dysentery. The development, birth, and death of Noah became a platform for conservationists and ethicists to critique the role of cloning in society and as a method to conserve species.

Created2013-07-26