This edited book covers all aspects of non-coding RNAs analysis. Non-coding RNAs are critical genetic regulators that have evolved as game-changers in modifying plant responses to stress conditions. The book explores potential relationships between different classes of non-coding RNAs and response of plants to biotic and abiotic stresses. The chapters are organized into three sections. Section 1 provides a basic understanding of the various aspects and roles of RNA silencing. Section 2 includes an overview of the molecular techniques and computational methodologies used for identification and analysis of small RNAs. Discovery timelines are also provided to highlight the significance of next-generation sequencing over time. Section 3 of the book covers the applications of RNAi techniques to improve crops and develop stress-tolerant varieties. This book compiles all available data and computational approaches to analyze non-coding RNAs. It describes the general characteristics, categorization, functional analysis, and advancements in RNAi and its role in developing stress-resilient plants. This book is helpful for students and researchers to understand the role of non-coding RNAs, which opens new insights into next-generation sequencing. It serves as excellent reading material for undergraduate and graduate students studying molecular biology, bioinformatics, agriculture, and environmental sciences.