Abstract kRAS is one of, if not the, most prevalent oncogenic aberrations identified to date, and has been well validated as a therapeutic target. It is either upregulated or mutationally activated in ∼30% of all cancers, including 60-90% of pancreatic cancer, and contributes to malignant transformation, tumorigenicity, and a corresponding poor prognosis. Our therapeutic approach to transcriptionally downregulate kRAS is innovate and promising because we are focused on ablating the oncogenic protein, rather than previously unsuccessful attempts focused on modulating its function. Numerous literature reports validate the downregulation of kRAS expression to as an anti-cancer approach, but there has yet to be a concerted effort in this area, primarily due to lack of a molecular target. This is where our findings are exceptionally pioneering, as we focus on globular higher order DNA (G4) formations within the core kRAS promoter as druggable targets with great promise. G4s occur in G/C-rich DNA and are made up of two or more stacked tetrads, formed by the Hoogsteen hydrogen bonding of four guanines, stabilized by monovalent cations. These G/C-rich regions preferentially cluster around the transcriptional start site throughout the genome peaking within 50 bp of the TSS, which is indicative of their regulatory role. Although not universally true, formation of G4s generally functions to silence transcription or translation and thus represents an attractive molecular target for kRAS. The proximal promoter of kRAS contains three separate putative G4-forming regions: near, mid, and far. We have revealed a previously undescribed within the “mid” region of the proximal kRAS promoter, which contains seven contiguous runs three or more continuous guanines. This region forms biologically functional, transcriptionally silencing, equilibrating G4s. This is in contrast to the previously described “near” G4, which we show to have little to no role in transcriptional regulation. These newly described mid-G4 structures have high potential as molecular targets for drug development. Citation Format: Jennifer Hockings, Tracy Ann Brooks. Identification of new, biologically relevant, G-quadruplexes formed within the critical promoter of kRAS as promising molecular targets. [abstract]. In: Proceedings of the AACR Special Conference on RAS Oncogenes: From Biology to Therapy; Feb 24-27, 2014; Lake Buena Vista, FL. Philadelphia (PA): AACR; Mol Cancer Res 2014;12(12 Suppl):Abstract nr A44. doi: 10.1158/1557-3125.RASONC14-A44