Matching Items (2)
Description
Cooperativity can be used to manipulate binding affinities of DNA biosensors – improving specificity without sacrificing sensitivity; examples include tentacle probes (TPs) and cooperative primers (CPs). This thesis body of work: (1) used TPs to develop a rapid, low-cost diagnostic for detecting the point mutation leading to Navajo Neurohepatopathy (NNH) and (2) used CPs to amplify a symmetric bowtie-barcoded origami with captured t-cell receptor (TCR) α and β mRNA of a single cell.
NNH (affecting 1-in-1600 Navajo babies) is a fatal genetic disorder often caused by 149G>A mutation and is characterized by brain damage and liver disease/failure. Phoenix Children’s Hospital currently uses gene sequencing to identify the 149G>A mutation. While this process is conclusive, there are limitations, as it requires both time (3-4 weeks) and money (>$700). Ultimately, these factors create barriers that can directly impact a patient’s quality of life. Assessment of the developed TP diagnostic, using genomic DNA derived from FFPE patient liver samples, suggests nearly 100% specificity and sensitivity while reducing cost to ~$250 (including cost of labor) and providing a diagnosis within 48 hours.
TCR specificity is dependent on V(D)J recombination as well as pairing of the αβ chains. Drs. Schoettle and Blattman have developed a solution in which a bowtie-barcoded origami strand nanostructure is transfected into individual cells of a heterogeneous cell population to capture and protect αβ mRNA. When PCR of the origami template is performed with Vα, X, Vβ, and Y primers, the α and β gene segments cannot be tied back to a barcode – and paired. Assessment of the developed CPs for PCR suggests correct individual amplification using (1) Va + Xcp and (2) Vβ + Ycp primers, whereas combination of all the primers (Va, Xcp, Vb, and Ycp) suggests hybridization of the Vα + Xcp and Vβ + Ycp products due to the origami target symmetry.
NNH (affecting 1-in-1600 Navajo babies) is a fatal genetic disorder often caused by 149G>A mutation and is characterized by brain damage and liver disease/failure. Phoenix Children’s Hospital currently uses gene sequencing to identify the 149G>A mutation. While this process is conclusive, there are limitations, as it requires both time (3-4 weeks) and money (>$700). Ultimately, these factors create barriers that can directly impact a patient’s quality of life. Assessment of the developed TP diagnostic, using genomic DNA derived from FFPE patient liver samples, suggests nearly 100% specificity and sensitivity while reducing cost to ~$250 (including cost of labor) and providing a diagnosis within 48 hours.
TCR specificity is dependent on V(D)J recombination as well as pairing of the αβ chains. Drs. Schoettle and Blattman have developed a solution in which a bowtie-barcoded origami strand nanostructure is transfected into individual cells of a heterogeneous cell population to capture and protect αβ mRNA. When PCR of the origami template is performed with Vα, X, Vβ, and Y primers, the α and β gene segments cannot be tied back to a barcode – and paired. Assessment of the developed CPs for PCR suggests correct individual amplification using (1) Va + Xcp and (2) Vβ + Ycp primers, whereas combination of all the primers (Va, Xcp, Vb, and Ycp) suggests hybridization of the Vα + Xcp and Vβ + Ycp products due to the origami target symmetry.
ContributorsDubois, Courtney Michelle (Author) / Caplan, Michael R (Thesis advisor) / Vernon, Brent (Committee member) / Carpentieri, David (Committee member) / Arizona State University (Publisher)
Created2018
Description
The Community Action Research Experiences (CARE) program collaborated with Singleton Moms, a local non-profit organization that provides financial, psychological, and social support services to single parents with cancer. The purpose of this action research project was to assess the volunteer program at Singleton Moms. Both past and present Singleton Moms' volunteers (N = 123; 87.0% female) completed an online survey assessing their motivation for volunteering and their satisfaction with the organization. A mixed ANOVA was conducted to identify the most important motivation and satisfaction domains and to see if the findings depended on whether the volunteers were current or past volunteers. For the motivation assessment, results indicated that the volunteers rate the cancer specific and moral/human kindness domains as the strongest reasons for motivating them to volunteer at Singleton Moms. In addition, results revealed that the social connection motivation domain was the only domain with differences between the ratings of the past and present volunteers. For the satisfaction assessment, results indicated that the volunteers rate the organizational climate domain as the most fulfilled area of satisfaction within the Singleton Moms' volunteer program. It was also revealed that there were no significant differences between the ratings of the past and present volunteers among all satisfaction domains. Both the quantitative and qualitative findings suggest that Singleton Moms' implications for action may include: 1) a volunteer database audit, 2) streamlining communications, 3) variability in volunteer times, and 4) bolstering volunteer motivation. Implementing some of these actions may help Singleton Moms increase volunteer motivation and satisfaction and thus create a more effective volunteer program. Ultimately, this may encourage volunteers to continue their services at Singleton Moms and thus help Singleton Moms expand their support programs and assist additional families.
ContributorsDubois, Courtney Michelle (Author) / Miller, Cindy (Thesis director) / Dumka, Larry (Committee member) / Harrington Bioengineering Program (Contributor) / Barrett, The Honors College (Contributor) / T. Denny Sanford School of Social and Family Dynamics (Contributor)
Created2016-05