RNAi and Small Regulatory RNAs in Stem Cells, Softcover reprint of the original 1st ed. 2017
Methods and Protocols
Methods in Molecular Biology Series, Vol. 1622

Coordinator: Zhang Baohong

Language: English

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This detailed collection provides an accessible compendium of up-to-date methods focused on the study of RNAi and small regulatory miRNAs in stem cells. Beginning with a brief introductory section, the volume continues by exploring methods and protocols for RNAi screening, transfection, and the knockdown of specific genes and pathways in several animal species, including humans and mice, recently developed methods for miRNA expression and functional analysis, as well as usage of CRISPR/Cas 9 to knockout an individual gene for functional studies. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, list of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Practical and authoritative, RNAi and Small Regulatory RNAs in Stem Cells: Methods and Protocols aims to accelerate progress in this crucial field by reducing the time required to decipher and put into practice procedures published in the literature.

Part I: Introduction

 

1. RNAi Technique in Stem Cell Research: Current Status and Future Perspectives

            Gang-Ming Zou

 

2. RNAi and MicroRNA-Mediated Gene Regulation in Stem Cells

            Alexander K. Murashov

 

Part II: RNA Interference

 

3. Forward RNAi Screens in Human Hematopoietic Stem Cells

            Roman Galeev, Christine Karlsson, Aurélie Baudet, and Jonas Larsson

 

4. Silencing of ATP11B by RNAi-Induced Changes in Neural Stem Cell Morphology

            Jiao Wang, Qian Wang, Fangfang Zhou, Dong Wang, and Tieqiao Wen

 

5. High-Efficiency Transfection of Glioblastoma Cells and a Simple Spheroid Migration Assay

            Carsten Hagemann, Diana Amend, Almuth F. Kessler, Thomas Linsenmann, Ralf-Ingo Ernestus, and Mario Löhr

 

6. Construction and Application of Random dsRNA Interference Library for Functional Genetic Screens in Embryonic Stem Cells

            Xiaoxing Cheng and Rui Jian

 

7. Efficient Depletion of Essential Gene Products for Loss-of-Function Studies in Embryonic Stem Cells

            Soizik Berlivet, Isabelle Hmitou, Hélène Picaud, 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. A Recessive Genetic Screen for Components of the RNA Interference Pathway Performed in Mouse Embryonic Stem Cells

            Melanie I. Trombly and Xiaozhong Wang

 

10. RNAi Knockdown of Ape1 Gene in the Differentiation of Mouse Embryonic Stem Cells

            Gang-Ming Zou, Jieqing Yu, Cynthia LeBron, and Yumei Fu

 

11. An Efficient Transfection Method for Differentiation and Cell Proliferation of Mouse Embryonic Stem Cells

            Jun-Yang Liou, Bor-Sheng Ko, and Tzu-Ching Chang

 

12. Implanting Glioblastoma Spheroids into Rat Brains and Monitoring Tumor Growth by MRI Volumetry

            Mario Löhr, Thomas Linsenmann, Anna Jawork, Almuth F. Kessler, Nils Timmermann, György A. Homola, Ralf-Ingo Ernestus, and Carsten Hagemann

 

13. RNAi-Based Techniques for the Analysis of Gene Function in Drosophila Germline Stem Cells

            Amelia J. Helms, Danielle S. Finger, Victoria L. Hardy, and Elizabeth T. Ables

 

14. In Vivo RNAi in the Drosophila Follicular Epithelium: Analysis of Stem Cell Maintenance, Proliferation, and Differentiation

            Veit Riechmann

 

15. A Phenotype-Based RNAi Screening for Ras-ERK/MAPK Signaling-Associated Stem Cell Regulators in C. elegans

            Myon-Hee Lee and Dong Suk Yoon

 

Part III: MicroRNAs

 

16. Engineering Human Mesenchymal Stem Cells to Release Adenosine Using miRNA Technology

            Gaoying Ren and Detlev Boison

 

17. Efficient Gene Knockdowns in Mouse Embryonic Stem Cells Using MicroRNA-Based shRNAs

            Jianlong Wang

 

18. Using Quantitative Real-Time PCR to Detect MicroRNA Expression Profile During Embryonic Stem Cell Differentiation

            Xiaoping Pan, Alexander K. Murashov, Edmund J. Stellwag, and Baohong Zhang

 

Part IV: CRISPR-Cas9 and Genome Editing

 

19. Genetic Tools for Self-Organizing Culture of Mouse Embryonic Stem Cells Via Small Regulatory RNA-Mediated Technologies, CRISPR/Cas9 and Inducible RNAi

            Nozomu Takata, Eriko Sakakura, Tetsushi Sakuma, and Takashi Yamamoto

 

20. CRISPR-Cas9-Mediated Gene Editing in Mouse Spermatogonial Stem Cells

            Yinghua Wang, Yifu Ding, and Jinsong Li

Includes cutting-edge methods and protocols for the study of the intersection of RNAi, small regulatory RNAs, and stem cell biology

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts