Molecular biology, which in a broad sense covers the manipulation of DNA and RNA, has become an integral part of most biological research programs. Research areas that utilize molecular biology techniques include cloning genes for expression of proteins in cell model systems, site-directed mutagenesis and tagging for cellular localization and functional studies, genotyping, RNA isolation and expression analysis, and the development of transgenic mice. The Molecular Biology Core was established at the University of North Carolina (UNC) Cystic Fibrosis Center in 1998 to provide molecular biology services and expertise to faculty who are often primarily trained in cell biology, physiology, or clinical sciences. The Core is currently an integral part of many of the ongoing projects described in the PPG providing expert guidance, technical support, and specialized equipment. All four projects within the PPG application will utilize Molecular Biology Core services. PPG investigators will require cloning of genes into retroviral, lentiviral and oocyte expression vectors for expression of a variety of tagged and/or modified proteins (CFTR, ENaC subunits, and nucleoside tranporters) in a variety of cell models, including well-differentiated human bronchial epithelial (HBE) (provided by Core C:Cell Culture). Secondly, RNA isolation and expression analysis services, including quantitative real-time PCR and development and testing of si/shRNA reagents, will be provided, along with more specialized services, such as RNA-sequencing and data analyses. Finally, the Molecular Biology Core will aid in the generation and utilization of new transgenic mouse models, i.e., Panxl and Slc17A9, to help identify important regulators of ainway surface liquid homeostasis. The Molecular Biology Core will effectively complement the expertise of project leaders and their staff and to allow them to focus on the experiments in their own personal areas of proficiency.

Public Health Relevance

The PPG focuses on pulmonary liquid homeostasis, attacking the problem from multiple vantage points in various cells, lung compartments, model systems. In as much as molecular biology can contribute to the successful completion of the goals of the grant, the Molecular Biology Core is highly relevant to the overall PPG. In this era of defining molecular mechanisms and links between muliple protein functions in complex model systems, molecular biology aspects of research projects will continue to be central and highly relevant.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Program Projects (P01)
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Heart, Lung, and Blood Initial Review Group (HLBP)
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University of North Carolina Chapel Hill
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Donoghue, Lauren J; Livraghi-Butrico, Alessandra; McFadden, Kathryn M et al. (2017) Identification of trans Protein QTL for Secreted Airway Mucins in Mice and a Causal Role for Bpifb1. Genetics 207:801-812
Sandefur, Conner I; Boucher, Richard C; Elston, Timothy C (2017) Mathematical model reveals role of nucleotide signaling in airway surface liquid homeostasis and its dysregulation in cystic fibrosis. Proc Natl Acad Sci U S A 114:E7272-E7281
Wang, Ling; Ariyarathna, Yamuna; Ming, Xin et al. (2017) A Novel Family of Small Molecules that Enhance the Intracellular Delivery and Pharmacological Effectiveness of Antisense and Splice Switching Oligonucleotides. ACS Chem Biol 12:1999-2007
Livraghi-Butrico, Alessandra; Grubb, Barbara R; Wilkinson, Kristen J et al. (2017) Contribution of mucus concentration and secreted mucins Muc5ac and Muc5b to the pathogenesis of muco-obstructive lung disease. Mucosal Immunol 10:395-407
Sesma, Juliana I; Weitzer, Clarissa D; Livraghi-Butrico, Alessandra et al. (2016) UDP-glucose promotes neutrophil recruitment in the lung. Purinergic Signal 12:627-635
Esther Jr, Charles R; Turkovic, Lidija; Rosenow, Tim et al. (2016) Metabolomic biomarkers predictive of early structural lung disease in cystic fibrosis. Eur Respir J 48:1612-1621
Dickey, Audrey S; Pineda, Victor V; Tsunemi, Taiji et al. (2016) PPAR-? is repressed in Huntington's disease, is required for normal neuronal function and can be targeted therapeutically. Nat Med 22:37-45
Yu, Dongfang; Davis, Richard M; Aita, Megumi et al. (2016) Characterization of Rat Meibomian Gland Ion and Fluid Transport. Invest Ophthalmol Vis Sci 57:2328-43
Shobair, Mahmoud; Dagliyan, Onur; Kota, Pradeep et al. (2016) Gain-of-Function Mutation W493R in the Epithelial Sodium Channel Allosterically Reconfigures Intersubunit Coupling. J Biol Chem 291:3682-92
Kirby, Brett S; Schwarzbaum, Pablo J; Lazarowski, Eduardo R et al. (2015) Liberation of ATP secondary to hemolysis is not mutually exclusive of regulated export. Blood 125:1844-5

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