Chronic pain often results from changes in nociceptors, specialized primary sensory neurons (PSNs) that detect painful stimuli. Following injury, nociceptors may become hyperactive, signaling pain even in the absence of harmful stimuli. Our single-cell sequencing data indicate that inflammation alters nociceptor activity patterns in response to neutral stimuli, which may contribute to the transition from acute to chronic pain. We aim to investigate how specific nociceptor subtypes change during this transition. At the same time, the set of genes essential for pain transduction remains poorly understood. Leveraging CRISPR-Cas9 technology, we will develop an in vivo multiplexed gene knockout assay to test the roles of multiple genes in nociceptor responses to pain stimuli. This requires building a robust analysis pipeline for gene transcript sequencing at both single-cell and bulk levels. By integrating these approaches, we will elucidate the mechanisms driving nociceptor changes in chronic pain and identify key genes involved in pain signaling, providing new insights into potential therapeutic targets for chronic pain conditions.