RNA interference (RNAi) is a promising molecular biology approach for combating viral diseases. In this study, we present an interference strategy targeting the viral βC1 gene to develop resistant okra lines against yellow vein mosaic virus (YVMV). The methodology involves several key steps, including total DNA isolation from infected plants, PCR amplification of the target gene, design of a hairpin RNAi construct harboring the βC1 gene of the β satellite of Begomovirus and utilizing the pRNAi-LIC (CD3-1285) vector. Successful transformation of the construct into the highly susceptible okra cv. Salkeerthi could be achieved using an in planta method. Additionally, the stem loop primers designed could amplify the target small interfering RNA (siRNA) through thermal cycling. Screening of transgenic lines is performed under containment conditions, involving the controlled release of infected vectors. This chapter provides detailed insights into the methodology and procedures for performing RNAi for the development of virus-resistant okra (Abelmoschus esculentus).