Abstract New Mexico is one of the most geographically and culturally diverse states and faces serious cancer health disparities among its poor, rural, and ethnically diverse populations of American Indians (AIs). They have lower cancer screening rates compared to other populations and are more likely to be diagnosed with a later stage of cancer but less likely to receive treatment (Li et al, 2003). In turn, AIs have the lowest five-year cancer survival rates compared to any ethnic/racial group in the US (Clegg, 2002; Edwards, 2005). Numerous barriers such as cultural beliefs in health and health care, fear, fatalism, mistrust, stigma and lack of culturally appropriate interventions contribute to low cancer screening rates and low cancer survivorship (Daley et al, 2012; Filippi et al, 2013; James et al, 2013). Zuni Health Initiative conducted a community-based participatory research project in collaborations with community stakeholders from the Zuni Pueblo to assess knowledge and perceptions about common cancers among Zuni adults. We used trained Community Health Representatives (CHRs) from Zuni along with Zuni undergraduate students who led six one-hour focus group sessions using structured questionnaire with probes designed to illicit information on knowledge and perceptions about cancer among Zuni adults. The focus groups were conducted among 51 participants from different age groups (20-29 yrs, n=19; 30-49 yrs, n=17; and 50 yrs and older, n=15) stratified by gender. Focus groups were conducted in English and Shiwi language and were audio recorded, with the Zuni students maintaining extensive notes on the focus group discussions. The Zuni CHRs transcribed the audio recordings, and developed a codebook based on the transcriptions and notes for the qualitative data analysis. Overall, the responses given by participants fell into three distinct categories including, (1) a general lack of knowledge, (2) the perception that the disease is uncontrollable in nature, and (3) general negative connotations. Although some participants acknowledged that cancer could come in many forms, there were many participants who were uncertain about the different factors that contribute to the disease, as well as specific outcomes associated with the disease. Many participants mentioned cancer being uncontrollable in nature, that cancer always comes back, and expressed that death is the inevitable outcome. Finally, several participants displayed general uncertainty and discomfort when discussing cancer. There was negative connotations when participants heard the word “cancer”, as well as construed negative implications about discussing personal experiences with cancer. Findings from this formative study provides evidence that can guide the development and testing of interventions aimed at enhancing knowledge about cancer and cancer-specific screening practices. Citation Format: Safia Safi, Donica Ghahate, Jeanette Bobelu, Angela Wandinger-Ness, Thomas Faber, Shiraz Mishra, Cheryl Willman, Vallabh (Raj) Shah. Assessing knowledge and perceptions about cancer among American Indians of Zuni Pueblo, New Mexico [abstract]. In: Proceedings of the Twelfth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2019 Sep 20-23; San Francisco, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl_2):Abstract nr B044.