Rnai and Microrna-Mediated Gene Regulation in Stem Cells: Methods, Protocols, and Applications

Part I: Introduction 1. RNAi in Stem Cells: Current Status and Future Perspectives Gang-Ming Zou 2. A Brief Introduction to RNAi and microRNAs in Stem Cells Alexander K. Murashov Part II: RNA Interference 3. Forward RNAi Screens in Human Stem Cells Christine Karlsson, Jonas Larsson, and Aur?lie Baudet 4. A Recessive Genetic Screen for Components of the RNA Interference Pathway in Mouse Embryonic Stem Cells Melanie I. Trombly and Xiaozhong Wang 5. Construction and Application of Random dsRNA Interference Library for Functional Genetic Screens in Embryonic Stem Cells Xiaoxing Cheng and Rui Jian 6. Establishing Efficient siRNA Knockdown in Stem Cells Using Fluorescent Oligonucleotides Stephen W. Chen and Steve K.W. Oh 7. Loss of Function Studies in Mouse Embryonic Stem Cells Using the pHYPER shRNA Plasmid Vector Soizik Berlivet, Martin Houlard, and Matthieu G?rard 8. Regulation and/or Repression of Cholinergic Differentiation of Murine Embryonic Stem Cells Using RNAi Directed Against Transcription Factor L3/Lhx8 Takayuki Manabe and Akio Wanaka 9. Silence of Rho-GDI? by RNAi Promotes the Differentiation of Neural Stem Cells Jiao Wang, Wei Lu, and Tieqiao Wen 10. RNAi Knockdown of Redox Signaling Protein Ape1 in the Differentiation of Mouse Embryonic Stem Cells Gang-Ming Zou, Cynthia LeBron, and Yumei Fu 11. Proteins Involved in Cell Migration from Glioblastoma Neurospheres Analyzed by Overexpression and siRNA-Mediated Knock-Down Carsten Hagemann, Harun M. Said, Michael Flentje, Klaus Roosen, and Giles Hamilton Vince 12. An Efficient Transfection Method for Mouse Embryonic Stem Cells Jun-Yang Liou, Bor-Sheng Ko, and Tzu-Ching Chang 13. Semi-Quantitative Analysis of Transient Single-Cell Gene Expression in Embryonic Stem Cells by Femtoinjection Mikako Saito and Hideaki Matsuoka Part III: microRNAs 14. Preparation and Analysis of microRNA Libraries using the Illumina Massively Parallel Sequencing Technology Ryan D. Morin, Yongjun Zhao, Anna-Liisa Prabhu, Noreen Dhalla, Helen McDonald, Pawan Pandoh, Angela Tam, Thomas Zeng, Martin Hirst, and Marco Marra 15. Assessing In Vivo MicroRNA Function in the Germline Stem Cells of the Drosophila Ovary Kin Chan and Hannele Ruohola-Baker 16. Monitoring microRNA Expression during Embryonic Stem Cell Differentiation Using Quantitative Real-Time PCR (qRT-PCR) Xiaoping Pan, Alexander K. Murashov, Edmund J. Stellwag, and Baohong Zhang 17. Engineering Human Mesenchymal Stem Cells to Release Adenosine Using miRNA Technology Gaoying Ren and Detlev Boison 18. Efficient Gene Knockdowns in Mouse Embryonic Stem Cells Using microRNA-Based shRNAs Jianlong Wang