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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
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Description
Immunology, the study of the immune system and its ability to distinguish self from non-self, is a rapidly advancing sector of molecular biology. Cancer, being host derived, provides a difficult challenge for immune cells to distinguish it from normal tissue. The historic treatment of cancer has had three main methods:

Immunology, the study of the immune system and its ability to distinguish self from non-self, is a rapidly advancing sector of molecular biology. Cancer, being host derived, provides a difficult challenge for immune cells to distinguish it from normal tissue. The historic treatment of cancer has had three main methods: radiation, chemotherapy, and surgery (1). Due to recent advancements in understanding the regulatory role of adaptive immunity against cancer, researchers have been attempting to engineer therapies to enhance patients’ immunities against their cancer. Immunotherapies, both passive and active, demonstrate potential for combating many diseases. Passive immunization provides temporary protection against a pathogen, whereas active immunization teaches the patient’s system to respond to the antigen independently, giving life-long immunity. Passive immunization, generally, is a much more expensive method of providing immunity and is commonly used in emergency situations. Anti-venom, for example, uses antibodies grown in lab to neutralize venom. Examples of active immunization are vaccines, which mimic the wild-type pathogen in a way that elicits an immune response, specifically naïve lymphocyte activation and maturation into memory lymphocytes. In terms of cancer therapy, both passive and active immunization are being tested for efficacy (2).
ContributorsMarquardt, Charles Andrew (Author) / Anderson, Karen S. (Thesis director) / Mason, Hugh S. (Committee member) / Lake, Douglas F. (Committee member) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
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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

During the twentieth and twenty-first centuries, Robert Paul Lanza studied embryonic stem cells, tissues,
and endangered species as chief scientific officer of Advanced Cell
Technology, Incorporated in Worcester, Massachusetts. Lanza's team cloned
the endangered species of gaur Bos gaurus.
Although the gaur did not survive long,

During the twentieth and twenty-first centuries, Robert Paul Lanza studied embryonic stem cells, tissues,
and endangered species as chief scientific officer of Advanced Cell
Technology, Incorporated in Worcester, Massachusetts. Lanza's team cloned
the endangered species of gaur Bos gaurus.
Although the gaur did not survive long, Lanza successfully cloned
another cow-like creature, called the banteng
(Bos
javanicus). Lanza also worked on cloning human embryos
to harvest stem cells, which could be used to treat dieases. While
previous techniques required the embryo's destruction, Lanza
developed a harvesting technique that does not destroy the embryo,
forestalling many ethical objections to human embryonic
research.

Created2015-02-11
Description

The Southern Gastric-Brooding Frog (Rheobatrachus silus) was an aquatic frog that lived in south-east Australia. In 2002, the International Union for Conservation of Nature Red List declared the frog extinct, although no wild specimens had been reported since 1981. As the common name alludes to, the R.
silus

The Southern Gastric-Brooding Frog (Rheobatrachus silus) was an aquatic frog that lived in south-east Australia. In 2002, the International Union for Conservation of Nature Red List declared the frog extinct, although no wild specimens had been reported since 1981. As the common name alludes to, the R.
silus was a gastric-brooder, meaning that the female's eggs developed inside of her stomach. Weeks after ingestion, juvenile frogs escape through the mother's mouth. Because no other observed species performs this reproductive behavior, in the early twenty-first century R. silus became a target of
the de-extinction movement that aims to resurrect extinct species. Researchers studied this frog's reproductive behavior and how the eggs and embryos escape digestion. Some scientists claimed that resurrecting this frog could result in future medical applications related to digestion and to reprogramming organ function, as during pregnancy, R. silus's stomach physiologically functioned as a uterus.

Created2015-01-26
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

In the early twentieth century US, Jean Paul Pratt and Edgar Allen conducted clinical experiments on women who had abnormal menstrual cycles. During the clinical tests, researchers injected the hormone estrogen into their patients to alleviate their menstrual ailments, which ranged from irregular cycles to natural menopause. The hormone estrogen

In the early twentieth century US, Jean Paul Pratt and Edgar Allen conducted clinical experiments on women who had abnormal menstrual cycles. During the clinical tests, researchers injected the hormone estrogen into their patients to alleviate their menstrual ailments, which ranged from irregular cycles to natural menopause. The hormone estrogen plays a prominent role in the menstrual cycle by signaling the tissue lining the uterus (endometrium) to thicken in preparation for possible pregnancy. In their clinical tests, Pratt and Allen showed that injecting estrogen into female human subjects restored their normal menstrual cycle, removed symptoms such as hot flashes, and caused uterine tissue to grow. The clinical tests conducted by Pratt and Allen provided experimental evidence and justification for the injection of isolated estrogen in women to alleviate, for a short amount of time, different menstrual problems, and it contributed to later hormone therapy research.

Created2017-04-06
Description
This project aims to tackle two perspectives: to design and express an enzyme that can perform single-molecule modifications for identification, and to determine the inclusion of the last adenosine in mature mRNAs within the metazoan, Caenorhabditis elegans. Starting with the first perspective, the enzymatic group that was utilized was methyltransferases.

This project aims to tackle two perspectives: to design and express an enzyme that can perform single-molecule modifications for identification, and to determine the inclusion of the last adenosine in mature mRNAs within the metazoan, Caenorhabditis elegans. Starting with the first perspective, the enzymatic group that was utilized was methyltransferases. Methyltransferases have gained great interest in biotechnology and academia due to their ability to make single-molecule modifications to a wide variety of biomolecules, ranging from proteins to RNA. Of these methyltransferases, the subset that has the greatest interest for this study are RNA methyltransferases. Of the known RNA methyltransferases, human METTL16 was chosen for this project, due to its ability to modify adenosines at the N6 position (m6A), specificity for its consensus motif, and its promise in chimeric enzymatic complexes. As a result of these properties, this study looks to design METTl16-based complexes for the purpose of identifying single nucleotides in RNA. The second perspective involves pre-mRNA cleavage and polyadenylation of the 3’ untranslated region (3’UTR). Cleavage of pre-mRNAs within C.elegans appears to prefer an adenosine, leading to the term “terminal adenosine” (terminal-A). Since RNA cleavage and polyadenylation is highly conserved across metazoans, we can utilize the model system, C. elegans, to apply our findings to humans. Utilizing METTL16’s ability to modify adenosines, it is theorized that it may be possible to modify the terminal-A in vivo within C. elegans. To confirm the functionality and utilization of METTL16, a novel methodology is currently being developed called the terminal adenosine methylation (TAM) assay. The TAM assay takes advantage of METTL16’s N-terminal RNA binding domain (RBD) and methyltransferase domain – called the “core” – to methylate the terminal adenosine of probe mRNA transcripts prior to cleavage in vivo. To determine if the adenosine is present within mature mRNAs, sequencing will determine if there is a m6A present, confirming that CPSF-3 cleaves either upstream or downstream of the terminal-A. Ultimately, this project focuses on designing METTL16 complexes for mRNA modification, testing the functionality of these constructs in vitro, and developing transgenic C. elegans strains to express the METTL16 complexes. The bioconjugation capabilities of RNA methyltransferases allow for concepts such as the TAM assay to be viable, as well as make way for future prospects of methyltransferases as a biotechnical tool.
ContributorsMurray, Jillian (Author) / Mangone, Marco (Thesis director) / Lapinaite, Audrone (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor) / School of Molecular Sciences (Contributor)
Created2024-05