mTOR: Methods and Protocols(821 Methods in Molecular Biology)

Since its discovery, the mammalian target of rapamycin (mTOR) has been shown to regulate many critical molecular processes in eukaryotes such as metabolism, growth, survival, aging, synaptic plasticity, memory, and immunity. In mTOR: Methods and Protocols expert researchers in the field detail many of the methods which are now commonly used to study mTOR. These include methods and techniques  used for the study of the mTOR pathway and potential therapeutic applications of mTOR inhibitors  such as, immunosuppressive and anticancer agents. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.   Authoritative and practical, mTOR: Methods and Protocols seeks to aid scientists in the further study of this crucially important molecule  ranging from the analysis of signal transduction events within a cell to the assessment of complex human diseases such as metabolic disorders or cancer.

Table of Contents:
Mammalian Target of Rapamycin: A Signaling Kinase for every Aspect of Cellular Life.-Biochemical and Pharmacological Inhibition of mTOR by Rapamycin and an ATP-Competitive mTOR Inhibitor.-Evaluation of the Nutrient-Sensing mTOR Pathway.-mTOR Activity Under Hypoxia.-Isolation of the mTOR Complexes by Affinity Purification.-An in vitro Assay for the Kinase Activity of mTOR Complex 2.-Overexpression or Downregulation of mTOR in Mammalian Cells.-Detection of Cytoplasmic and Nuclear Functions of mTOR by Fractionation.-Evaluation of Rapamycin-Induced Cell Death.-Evaluation of mTOR-Regulated mRNA Translation.-A Genome-wide RNAi Screen for Polypeptides that Alter rpS6 Phosphorylation.-Immunohistochemical Analysis of mTOR activity in Tissues.-Assessing Cell Size and Cell Cycle Regulation in Cells With Altered TOR Activity.-Quantitative Visualization of Autophagy Induction by mTOR Inhibitors.-The in vivo Evaluation of Active-Site TOR Inhibitors (asTORi) in Models of BCR-ABL+ Leukemia.-Inducible raptor and rictor Knockout Mouse Embryonic Fibroblasts.-Expanding Human T Regulatory Cells with the mTOR-Inhibitor Rapamycin.-Rapamycin Induced Enhancement of Vaccine Efficacy in Mice.-Utilizing a Retroviral RNAi System to Investigate in vivo mTOR Functions in T Cells.-Exploring Functional in Vivo Consequences of the Selective Genetic Ablation of mTOR Signaling in T Helper Lymphocytes.-Evaluating the Therapeutic Potential of mTOR Inhibitors Using Mouse Genetic.- Inhibition of PI3K-Akt-mTOR Signaling in Glioblastoma by mTORC1/2 Inhibitors.-Assessing the Function of mTOR in Human Embryonic Stem Cells.-Video-EEG Monitoring Methods for Characterizing Rodent Models of Tuberous Sclerosis and Epilepsy.-A Genetic Model to Dissect the Role of Tsc-mTORC1 in Neuronal Cultures.-Tissue-Specific Ablation of Tsc1 in Pancreatic Beta-Cells.-A Mouse Model of Diet-Induced Obesity and Insulin Resistance.-Rapamycin as Immunosuppressant in Murine Transplantation Model.-Development of ATP-Competitive mTOR Inhibitors.