Sustainable agriculture is severely affected by insect as well as pest attack. The manipulation of numerous genes to control pests by RNAi involves post-transcriptional gene silencing that is sequence-specific. This chapter is the first to describe the use of hairpin RNAi in tobacco plants as a novel approach to target insect pests and the potential impact this method could have on future pest management. Here we describe the generation of hpRNA constructs in tobacco for controlling insect pests. First, it describes how to make and test hpRNA constructs aimed at essential pest genes such as those required for development or survival. In this part of our study, the chapter on RNAi design explains in depth the procedures to create these RNAi vectors using cloning strategies in vector structure. It also deals with the use of Agrobacterium-mediated transformation for both tobacco plants and selection of transgenic plants. We look at approaches for measuring the impact of RNAi in these plants, such as gene expression analysis and evaluation of host resistance in laboratory or field conditions. This chapter is a comprehensive guide by giving an in-depth view on the steps from construct designing to in field applications; thus it will serve as reference for all future workers of RNAi technology looking towards effective and eco-friendly ways of pest management in agriculture.