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
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.