DNA, the blue print of life, is densely wrapped around histone proteins to form chromatin. Chromatin remodeling ATPases unwind histone-DNA interactions to facilitate DNA transcription, modification, and repair. Four genes involved in chromatin remodeling, namely, imitation SWI (iswi), chromodomain-helicase-DNA-binding protein 1 (chd-1), DNA helicase INO80 (ino80) and mi-2 were silenced through the injection of dsRNA, and phenotypes were assessed in bed bugs. Bed bugs were injected with 0.2 μg dsRNA per insect between the last thoracic segment and first abdominal segment using a fine capillary tube fitted to a nanoinjector. We observed a significant reduction in reproductive potential with all the genes tested, suggesting the essential function of chromatin remodeling ATPases in many cellular processes including egg-laying and egg-hatching. Knockdown of mi-2 and iswi completely inhibited oviposition over time. Real-time quantitative polymerase chain reaction confirmed significant knockdown of targeted mRNAs for at least 30 d, which supports persistence of RNAi in bed bugs. In addition, we observed a significant depletion of targeted transcripts in eggs laid by bed bugs injected with dsRNAs specific to chromatin remodeling ATPases. This study demonstrates the importance of chromatin remodeling ATPase in bed bug reproduction.