We will make fully operational a continuing pilot collaboration between the labs of Dr. Jessica P. Houston (NMSU) and Dr. Roger Brent (FHCRC). Our proposed work builds on complementary strengths in the two labs: instrumentation engineering and signal analysis in the Houston lab, protein engineering and genetics in the Brent lab. In particular, the work builds on """"""""frequency domain"""""""" methods to determine fluorescence lifetimes in microscopy, and on significantly more advanced developments in """"""""frequency domain"""""""" methods for flow cytometry, for which Dr. Houston is emerging as a leader. Dysfunctions in cell signaling leading to inappropriate cell proliferation contribute to most cancers. Although much is known about cell signaling, key questions, including the causes and consequences of cell-to-cell variation in signaling and response, remain unanswered. Here, we will use fluorescent lifetime methods to extend the power of single cell assays to quantify key events in cell signaling in yeast (the """"""""development platform"""""""") and in mammalian cells. These measurements depend on the use of Green Fluorescent Protein (GFP) derivatives fused to the proteins that make up the different cell signaling pathways to quantify events such as recruitment of a signaling complex to the cell membrane. During the next three years, we will develop our methods, to address these questions and to allow wider applications. In particular, we will continue to improve our instrumentation, use it to develop better GFP derivatives and signaling reporters, and use the improved methods to understand the causes of cell-to-cell variation in a yeast signaling system. We will also use these methods to develop high-signal reporters that allow quantification of cell signaling in clonal, mammalian cell lines using flow cytometry. Successful work will merit future funding. It should also increase the visibility of the Houston lab and help recruit new students and researchers from underrepresented ethnic minorities into an active area of international scientific research.
| Grandori, Carla; Kemp, Christopher J (2018) Personalized Cancer Models for Target Discovery and Precision Medicine. Trends Cancer 4:634-642 |
| Ortega, Sigolène; McAlvain, Megan Stamey; Briant, Katherine J et al. (2018) Perspectives of Community Advisory Board Members in a Community-Academic Partnership. J Health Care Poor Underserved 29:1529-1543 |
| Sanchez, N S; Quinn, K E; Ashley, A K et al. (2018) In the ovine pituitary, CXCR4 is localized in gonadotropes and somatotropes and increases with elevated serum progesterone. Domest Anim Endocrinol 62:88-97 |
| Molina, Yamile; Briant, Katherine J; Sanchez, Janeth I et al. (2018) Knowledge and social engagement change in intention to be screened for colorectal cancer. Ethn Health 23:461-479 |
| Lui, Goldie Y L; Grandori, Carla; Kemp, Christopher J (2018) CDK12: an emerging therapeutic target for cancer. J Clin Pathol 71:957-962 |
| Gurley, Kay E; Ashley, Amanda K; Moser, Russell D et al. (2017) Synergy between Prkdc and Trp53 regulates stem cell proliferation and GI-ARS after irradiation. Cell Death Differ 24:1853-1860 |
| Tham, Heidi M; Hohl, Sarah; Copeland, Wade et al. (2017) Enhancing Biospecimen Knowledge Among Health Care Providers and Representatives From Community Organizations. Health Promot Pract 18:715-725 |
| Pauli, Chantal; Hopkins, Benjamin D; Prandi, Davide et al. (2017) Personalized In Vitro and In Vivo Cancer Models to Guide Precision Medicine. Cancer Discov 7:462-477 |
| Ornelas, India J; Deschenie, Desiree; Jim, Jesse et al. (2017) Yéego Gardening! A Community Garden Intervention to Promote Health on the Navajo Nation. Prog Community Health Partnersh 11:417-425 |
| Sands, Bryan; Brent, Roger (2016) Overview of post Cohen-Boyer methods for single segment cloning and for multisegment DNA assembly. Curr Protoc Mol Biol 113:3.26.1-3.26.20 |
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