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Cover image for RNAi and microRNA-mediated gene regulation in stem cells : methods, protocols, and applications
Title:
RNAi and microRNA-mediated gene regulation in stem cells : methods, protocols, and applications
Series:
Methods in molecular biology ; 650

Springer protocols
Edition:
1st ed.
Publication Information:
New York : Springer, 2010.
Physical Description:
xi, 261 p. : ill. (some col.) ; 27 cm.
ISBN:
9781607617686

9781607617693

Available:*

Library
Item Barcode
Call Number
Material Type
Item Category 1
Status
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30000010274341 QH450.25 R53 2010 Open Access Book Book
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On Order

Summary

Summary

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.


Table of Contents

Gang-Ming ZouAlexander K. MurashovChristine Karlsson and Jonas Larsson and Aurélie BaudetMelanie I. Trombly and Xiaozhong WangXiaoxing Cheng and Rui JianStephen W. Chen and Steve K.W. OhSoizik Berlivet and Martin Houlard and Matthieu GérardTakayuki Manabe and Akio WanakaJiao Wang and Wei Lu and Tieqiao WenGang-Ming Zou and Cynthia LeBron and Yumei FuCarsten Hagemann and Harun M. Said and Michael Flentje and Klaus Roosen and Giles Hamilton VinceJun-Yang Liou and Bor-Sheng Ko and Tzu-Ching ChangMikako Saito and Hideaki MatsuokaRyan D. Morin and Yongjun Zhao and Anna-Liisa Prabhu and Noreen Dhalla and Helen McDonald and Pawan Pandoh and Angela Tam and Thomas Zeng and Martin Hirst and Marco MarraKin Chan and Hannele Ruohola-BakerXiaoping Pan and Alexander K. Murashov and Edmund J. Stellwag and Baohong ZhangGaoying Ren and Detlev BoisonJianlong Wang
Prefacep. v
Contributorsp. ix
Section I Introduction
1 RNAi in Stem Cells: Current Status and Future Perspectivesp. 3
2 A Brief Introduction to RNAi and MicroRNAs in Stem Cellsp. 15
Section II RNA Interference
3 Forward RNAi Screens in Human Stem Cellsp. 29
4 A Recessive Genetic Screen for Components of the RNA Interference Pathway in Mouse Embryonic Stem Cellsp. 45
5 Construction and Application of Random dsRNA Interference Library for Functional Genetic Screens in Embryonic Stem Cellsp. 65
6 Establishing Efficient siRNA Knockdown in Stem Cells Using Fluorescent Oligonucleotidesp. 75
7 Loss-of-Function Studies in Mouse Embryonic Stem Cells Using the pHYPER shRNA Plasmid Vectorp. 85
8 Regulation and/or Repression of Cholinergic Differentiation of Murine Embryonic Stem Cells Using RNAi Directed Against Transcription Factor L3/Lhx8p. 101
9 Silencing of Rho-GDIy by RNAi Promotes the Differentiation of Neural Stem Cellsp. 111
10 RNAi Knockdown of Redox Signaling Protein Apel in the Differentiation of Mouse Embryonic Stem Cellsp. 121
11 Proteins Involved in Cell Migration from Glioblastoma Neurospheres Analyzed by Overexpression and siRNA-Mediated Knock-Downp. 129
12 An Efficient Transfection Method for Mouse Embryonic Stem Cellsp. 145
13 Semi-quantitative Analysis of Transient Single-Cell Gene Expression in Embryonic Stem Cells by Femtoinjectionp. 155
Section III MicroRNAs
14 Preparation and Analysis of MicroRNA Libraries Using the Illumina Massively Parallel Sequencing Technologyp. 173
15 Assessing In Vivo MicroRNA Function in the Germline Stem Cells of the Drosophila Ovaryp. 201
16 Monitoring MicroRNA Expression During Embryonic Stem-Cell Differentiation Using Quantitative Real-Time PCR (qRT-PCR)p. 213
17 Engineering Human Mesenchymal Stem Cells to Release Adenosine Using miRNA Technologyp. 225
18 Efficient Gene Knockdowns in Mouse Embryonic Stem Cells Using MicroRNA-Based shRNAsp. 241
Subject Indexp. 257
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