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RNAi and microRNA-Mediated Gene Regulation in Stem Cells
Methods, Protocols, and Applications
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Main description:

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 (TM) 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.


Contents:

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 Aurelie 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 Gerard

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


PRODUCT DETAILS

ISBN-13: 9781607617686
Publisher: Springer (Humana Press Inc.)
Publication date: August, 2010
Pages: 251
Weight: 784g
Availability: Available
Subcategories: Biochemistry, Genetics
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