1;RNA Technologies and Their Applications;3 1.1;Preface;5 1.2;Contents;9 1.3;Contributors;11 1.4;The Key Features of RNA Silencing;17 1.4.1;1 Introduction;18 1.4.2;2 RNA Silencing Effector as a Two-Component System;21 1.4.3;3 Small RNA Biogenesis;22 1.4.3.1;3.1 miRNAs and siRNAs;22 1.4.3.2;3.2 Dicer-Independent Pathways;24 1.4.3.2.1;3.2.1 piRNAs;24 1.4.3.2.2;3.2.2 Secondary siRNAs in Worm;26 1.4.3.3;3.3 Endo-siRNAs;27 1.4.4;4 RISC Loading, Sorting, and Target-Sensing of Small RNAs;28 1.4.4.1;4.1 Sorting by Precursor Structures;30 1.4.4.2;4.2 Sorting by the 5 Ends;31 1.4.4.3;4.3 Sorting by Dicer Processing Polarity;31 1.4.4.4;4.4 Target-Sensing;32 1.4.5;5 Safeguards for RNA Silencing Pathways;33 1.4.6;6 Effector Modes of RNA Silencing Pathways;34 1.4.7;7 Regulations of RNA Silencing Pathways;35 1.4.7.1;7.1 Processing;35 1.4.7.2;7.2 Modification;37 1.4.7.3;7.3 RISC Activity;37 1.4.8;8 Perspective;38 1.4.9;References;39 1.5;Selected Strategies for the Delivery of siRNA In Vitro and In Vivo;45 1.5.1;1 Introduction;47 1.5.2;2 Mechanism of RNA Interference;48 1.5.3;3 Naked Delivery of siRNA In Vitro;48 1.5.3.1;3.1 Cellular Uptake of Naked Nucleic Acids;48 1.5.3.2;3.2 The Phosphorothioate-Stimulated Cellular Delivery of siRNA;50 1.5.3.3;3.3 The siRNA-Peptide Conjugate Approach;51 1.5.3.4;3.4 Intracellular Release of siRNA: A Major Hurdle;54 1.5.4;4 CPP-Mediated siRNA Delivery;55 1.5.4.1;4.1 Cell-Penetrating Peptides;55 1.5.4.2;4.2 Selected Examples of CPP-Mediated siRNA Delivery;57 1.5.5;5 Selected Examples of siRNA Delivery In Vivo;63 1.5.6;6 Conclusions and Future Prospects;66 1.5.7;References;67 1.6;RNAi Suppression and Its Application;75 1.6.1;1 RNA Interference;77 1.6.2;2 RNAi-Directed Viral Immunity;81 1.6.3;3 Identification and Function Characterization of Viral RNAi Suppressors;84 1.6.3.1;3.1 Agrobacterium-Mediated Transient Suppression Assay;85 1.6.3.2;3.2 Reversal of Transgene-Induced Gene Silencing;86 1.6.3.3;3.3 Grafting Assay;86 1.6.3.4;3.4 Replication Rescue of Mutant Viruses Defective in RNAi Suppression;86 1.6.4;4 Function Mechanism of Viral RNAi Suppressors;88 1.6.4.1;4.1 Viral Suppressors that Bind Viral dsRNA to Interfere with viRNA Biogenesis;89 1.6.4.2;4.2 Viral Suppressors that Target Virus-Derived siRNA for RNAi Suppression;91 1.6.4.3;4.3 Viral Suppressors that Target RNAi Effectors for Suppression;93 1.6.4.4;4.4 Viral Suppressors that Suppress Systemic RNAi;95 1.6.5;5 RNAi Suppressors of Nonviral Origin;96 1.6.5.1;5.1 Suppression of RNAi by Bacterial Pathogens;96 1.6.5.2;5.2 Cellular RNAi Suppressors;98 1.6.6;6 Biotechnological Application of RNAi Suppressors;99 1.6.6.1;6.1 Enhance Gene Expression for Rapid Function Analysis and Mass Production of Valuable Protein;99 1.6.6.2;6.2 Serve as Molecular Tools to Probe Various RNAi-Directed Functions;100 1.6.7;Appendix;101 1.6.8;References;103 1.7;Strategies to Prevent siRNA-Triggered Cellular Toxicity;109 1.7.1;1 Introduction;110 1.7.2;2 Cellular Sensors of siRNA-Triggered Innate Immune Response;112 1.7.2.1;2.1 TLR-Mediated Innate Immunity;113 1.7.2.2;2.2 Non-TLR-Mediated Innate Immunity;113 1.7.3;3 Cellular Sequels After siRNA-Triggered Innate Immune System Activation;114 1.7.4;4 Overcoming Synthetic siRNA-Triggered Innate Immune Response;115 1.7.5;5 Overcoming shRNA-Triggered Innate Immune Response;116 1.7.6;6 shRNA-Mediated Disruption of the Endogenous miRNA Machinery;118 1.7.7;7 Conclusions;119 1.7.8;References;119 1.8;RNAi in Malignant Brain Tumors: Relevance to Molecular and Translational Research;123 1.8.1;1 Introduction;124 1.8.1.1;1.1 Diagnostic Characteristics of Diffuse Astrocytic Tumor;124 1.8.1.2;1.2 Clinical Course;125 1.8.1.2.1;1.2.1 Symptoms;125 1.8.1.2.2;1.2.2 Prognosis;126 1.8.1.3;1.3 Obstacles in Treatment of Diffuse Astrocytic Tumors;126 1.8.2;2 RNAi for Glioma: from Bench to Clinic;127 1.8.2.1;2.1 Preclinical Studies;128 1.8.2.2;2.2 Target Genes for Silencing;129 1.8.2.3;2.3 Problems in Clinical Translation;130 1.8.2.3.1;2.3.1 Specificity;131 1.