RNAi and Small Regulatory RNAs in Stem Cells

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.

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

Inhalt

RNAi Technique in Stem Cell Research: Current Status and Future Perspectives.- RNAi and MicroRNA-Mediated Gene Regulation in Stem Cells.- Forward RNAi Screens in Human Hematopoietic Stem Cells.- Silencing of ATP11B by RNAi-Induced Changes in Neural Stem Cell Morphology.- High-Efficiency Transfection of Glioblastoma Cells and a Simple Spheroid Migration Assay.- Construction and Application of Random dsRNA Interference Library for Functional Genetic Screens in Embryonic Stem Cells.- Efficient Depletion of Essential Gene Products for Loss-of-Function Studies in Embryonic Stem Cells.- Regulation and/or Repression of Cholinergic Differentiation of Murine Embryonic Stem Cells Using RNAi Directed Against Transcription Factor L3/Lhx8.- A Recessive Genetic Screen for Components of the RNA Interference Pathway Performed in Mouse Embryonic Stem Cells.- RNAi Knockdown of Ape1 Gene in the Differentiation of Mouse Embryonic Stem Cells.- An Efficient Transfection Method for Differentiation and Cell Proliferation of Mouse Embryonic Stem Cells.- Implanting Glioblastoma Spheroids into Rat Brains and Monitoring Tumor Growth by MRI Volumetry.- RNAi-Based Techniques for the Analysis of Gene Function in Drosophila Germline Stem Cells.- In Vivo RNAi in the Drosophila Follicular Epithelium: Analysis of Stem Cell Maintenance, Proliferation, and Differentiation.- A Phenotype-Based RNAi Screening for Ras-ERK/MAPK Signaling-Associated Stem Cell Regulators in C. elegans.- Engineering Human Mesenchymal Stem Cells to Release Adenosine Using miRNA Technology.- Efficient Gene Knockdowns in Mouse Embryonic Stem Cells Using MicroRNA-Based shRNAs.- Using Quantitative Real-Time PCR to Detect MicroRNA Expression Profile During Embryonic Stem Cell Differentiation.- Genetic Tools for Self-Organizing Culture of Mouse Embryonic Stem Cells Via Small Regulatory RNA-Mediated Technologies, CRISPR/Cas9 and Inducible RNAi.- CRISPR-Cas9-Mediated Gene Editing in Mouse Spermatogonial Stem Cells.