NSF Award
2015806
The central goal of this project is to provide a better understanding of the molecular mechanisms that ensure that the genetic material within living cells is used properly and efficiently. Proper utilization of genetic material is crucial for all aspects of life, ranging from the ability of single-celled organisms to sense and respond to their surroundings to processes that promote the development of a newborn human being from a fertilized egg. Thus, this research will contribute to our understanding of fundamental questions of widespread biological importance. The project will include participation of a large number of undergraduate students as key scientific contributors. Twelve students will be carrying out experiments in the PI?s laboratory: these students will be mentored by the PI in various aspects of the scientific process, including experimental design and execution, presentation of research at scientific meetings, and critical evaluation of scientific literature. These students will also have the opportunity to develop skills required for effective communication of scientific concepts to the general public. Approximately 100 additional students will carry out part of this project in the context of laboratory courses: these students will learn cutting edge experimental approaches and will be engaged in discussions related to the uses and ethical considerations of current gene editing technologies. <br/><br/>Within eukaryotic cells, most of the DNA resides within the cell nucleus and is highly compacted through association with a number of proteins to form a structure known as chromatin, the basic unit of which is referred to as the nucleosome. Whereas nucleosomes are of critical importance to cells, they also represent physical obstacles to cellular factors whose functions require access to DNA. During gene transcription, a complex known as FACT is recruited to genes to assist with the disassembly of nucleosomes to facilitate RNA Polymerase II (Pol II) transcription, as well as to promote nucleosome re-assembly in the wake of Pol II passage. Previous work from the PI?s laboratory using the Saccharomyces cerevisiae model system identified a region on the side of the nucleosome, referred to as the ISGI region, required for proper dissociation of FACT from several genes following transcription. In this project, the PI?s team will use genome-wide approaches to study the role of the ISGI region in controlling FACT-chromatin interactions across the entire yeast genome and in promoting other chromosomal processes. Experiments will also be carried out to identify and characterize additional nucleosomal regions and proteins that influence FACT?s ability to interact with genes. Finally, a CRISPR-based approach will be used to simultaneously identify proteins that assist Pol II in removing non-nucleosomal obstacles in its path and to identify factors that promote Cas9-gRNA localization to target genomic sites. Since the factors and processes studied in this project are highly conserved across evolution, findings stemming from this research will shed new light on processes relevant to all eukaryotes, including humans.<br/><br/>This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.
