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Full-Text Articles in Life Sciences

Crispr/Cas9 Whole Genome Screens Reveal Novel Regulators Of Endocytic Processes In Macrophages, Jared Wollman Jan 2020

Crispr/Cas9 Whole Genome Screens Reveal Novel Regulators Of Endocytic Processes In Macrophages, Jared Wollman

Electronic Theses and Dissertations

Macrophages maintain tissue homeostasis by identifying and eliminating threats within their tissue microenvironment. Pattern recognition receptors allow macrophages to recognize and internalize specific ligands while macropinocytosis allows the internalization of all extracellular solutes from their environment. Without pattern recognition receptors, pathogens may grow unchecked if they cannot be detected, and without macropinocytosis, macrophages struggle to detect and move around their environment. This thesis presents the results of CRISPR/Cas9 whole-genome screens used to identify the regulators of both endocytosis (Chapter 2) and macropinocytosis (Chapter 3). In Chapter 2, we report genes regulating dextran uptake in primary murine macrophages and reveal Mrc1-mediated …


The Contributions Of Fc Gamma Receptors And Macropinocytosis To The Internalization, Sorting And Clearance Of Antibody Coated Nanovesicles In Macrophages, George Opoku-Kusi Jr. Jan 2018

The Contributions Of Fc Gamma Receptors And Macropinocytosis To The Internalization, Sorting And Clearance Of Antibody Coated Nanovesicles In Macrophages, George Opoku-Kusi Jr.

Electronic Theses and Dissertations

Macrophages are tissue-resident phagocytes that play critical roles in immune response and tissue homeostasis. They have a tremendous capacity to internalize objects of various sizes ranging from nanoscale viral particles to micron sized bacteria and tumor cells. This phenomenon, termed phagocytosis for the uptake of large particles, or endocytosis for the uptake of small particles, is integral to the immune response in multicellular organisms. Macrophages express FcγRs, which are tyrosine kinase receptors that bind IgG on an opsonized target. Binding of IgG Fc domain to the extracellular domain of an FcγR triggers signaling cascades that coordinate internalization of the opsonized …


Macropinosome Maturation Is A Clathrin Dependent Process In Bone Marrow Macrophages, Susmita Poudel Jan 2017

Macropinosome Maturation Is A Clathrin Dependent Process In Bone Marrow Macrophages, Susmita Poudel

Electronic Theses and Dissertations

Macrophages nonspecifically take up extracellular fluids, solutes and macromolecules by macropinocytosis. Understanding the mechanisms of macropinosome maturation will inform the study of lipid uptake, viral entry, antigen processing and presentation, as well as regulation of cell growth. Colony stimulating factor-1 receptor (CSF-1R) is internalized by small vesicle endocytosis, trafficked to nascent macropinosomes and degraded. These CSF-1R positive macropinosomes mature through a sequence similar to endosomes, progressing from EEA1 and Rab5 to Rab7 positive vesicles before fusing with lysosomes. Here we report the assembly of clathrin on internalized macropinosomes shown both by live-cell microscopy of cells expressing clathrin light chain-yellow fluorescent …


Silencing Seed Dormancy Genes To Mitigate Risk Of Transgene Flow To Weedy Rice, Alexander Wireko Kena Jan 2017

Silencing Seed Dormancy Genes To Mitigate Risk Of Transgene Flow To Weedy Rice, Alexander Wireko Kena

Electronic Theses and Dissertations

The flow of fitness-enhancing transgenes from genetically modified crops into wild/weed relatives may cause serious ecological and economic consequences. Seed dormancy (SD) is a key adaptive trait that distributes germination over time, resulting in weed persistence in agroecosystems. Thus, silencing major genes controlling SD would reduce the adaptive fitness of weeds. SD-enhancing genes cloned from weedy rice include SD7-1, SD7-2, SD12a, SD12b, and SD12c. The goal of this study was to develop a transgenic mitigation (TM) strategy using SD gene-silencing structures as mitigating factors to reduce the risk of transgene flow to wild/weed populations. TM vector constructs consisted of the …