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

ISBN-13: 9781493957491 / Angielski / Miękka / 2016 / 261 str.

Baohong Zhang; Edmund J. Stellwag

Opis

This cutting-edge text will be a valuable resource for scientists and aspiring graduate students interested in the intersection of RNAi, miRNA, and stem cell molecular biology and exciting areas of research such as regenerative medicine and cancer.

Spis treści

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 Aurélie 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 Gérard 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

Blurb

Recent stem cell research has revealed that miRNA and RNAi-mediated gene regulation is one of the vital determinates controlling the state of cell differentiation, with the small RNAs serving as key elements involved in regulatory network control of pluripotent cell fate determination. In RNAi and microRNA-Mediated Gene Regulation in Stem Cells: Methods, Protocols, and Applications, expert authors from laboratories across the globe contribute an accessible compendium of up-to-date, proven methods focused on the study of the titular topic. Divided into three sections, the book first gives a brief introduction to RNAi and miRNAs in stem cells, with a focus on the current status of research and future perspectives, then it continues with detailed methods and protocols for RNAi screening, transfection, and the knockdown of specific genes and pathways in several animal species, including humans and mice, concluding with a section on recently developed methods for identification of miRNAs, including a general protocol for preparation and analysis of miRNA libraries for deep sequencing, knock down of a specific gene using miRNA-based shRNA, and miRNA expression analysis using qRT-PCR. Written in the highly successful Methods in Molecular Biology™ series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes highlighting tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, RNAi and microRNA-Mediated Gene Regulation in Stem Cells: Methods, Protocols, and Applications serves as a valuable resource for scientists and aspiring graduate students interested in the intersection of RNAi, miRNA, and stem cell molecular biology and the exciting areas of medicine, including regenerative medicine, aging, cancer, and neurological disorders, that can be advanced through this expanding area of research.