Abstract The current landscape of druggable disease targets represents only ~10% of the human genome. Among those targets that are frequently mutated in cancer yet remain undruggable by conventional small molecules are the Ras family of GTPases. Somatic mutations in KRAS occur in 35% of colorectal cancers, 16% of lung cancers and 61% of pancreatic cancers. Common KRAS mutations lock the enzyme in the active GTP-bound state, which leads to constitutive activation of downstream effector modules such as MAPK, PI3K, Ral, Rac and Rho, which promote tumor growth, proliferation and survival. RNAi interference allows for the ablation of any gene product through the engagement of the endogenous microRNA machinery. The successful delivery of siRNA against KRAS or downstream effectors to tumors would represent a novel alternative treatment for KRASmut cancers, which currently lack strong clinical options. We have thus developed an RNAi therapy that employs 1) a cyclodextrin-based nanoparticle with high siRNA carrying capacity and tumor-targeting modules; and 2) highly potent “Sensor” siRNAs that can be used at low dose to efficiently ablate gene targets while avoiding off-target effects. We have assembled a library of potent Sensor siRNAs, which are identified using a massively parallel biological assay. These siRNA sequences are screened in high-throughput in KRASmut cell lines for negative effects on viability and disruption of cellular signaling pathways (ViBE, Bioscale). To subsequently assess the ability of these personalized nanoparticle-siRNA treatments to inhibit the growth of KRASmut tumors, we have developed an in vivo screening platform to monitor nanoparticle-siRNA delivery and payload efficacy at varying siRNA doses. Successful delivery of siRNAs to solid tumors has been one of the major hurdles to the advancement of RNAi therapy. Our fluorescent reporter system thus allows for the careful tracking of nanoparticle delivery and siRNA payload activity in real time using optical imaging. Additionally, the effect of different nanoparticle-siRNA treatments on transduced tumor cells can be measured non-invasively in real time. We demonstrate that delivery of low dose Sensor siRNAs targeting KRAS and other effector genes to KRASmut xenograft tumors significantly impedes tumor growth by inhibition of MAPK signaling and induction of apoptosis. The highly customizable nature of the siRNA payload offers a promising and universal platform to access previously intractable targets such as KRAS and personalize treatment for various oncogene-addicted tumors. Citation Format: Tina L. Yuan, Chih-Shia Lee, Christof Fellmann, Cayde Ritchie, Changwoo Lee, Colin Merrifield, Thomas Schluep, Scott W. Lowe, Ji Luo, Frank McCormick. Nanoparticle-based RNAi therapy for the delivery of personalized siRNA payloads to KRAS-driven tumors. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr LB-94. doi:10.1158/1538-7445.AM2013-LB-94