ABSTRACT. Elk prion protein (PrP C ) has been confirmed to be capable of rendering rabbit epithelial RK13 cells permissive to temporal infection by chronic wasting disease (CWD) prions. The present study satisfactorily generated persistently CWD prion-affected RK13 cells (RKC1-11) using elk PrP C expressing cells (elkRK13) that were generated via the lentiviral expression system with high efficiency. The elkRK13 cells have been shown to be permissive to accumulation of abnormal isoforms of prion protein (PrP Sc ) resulting from CWD prions up to 97 serial passages thus far. This novel prion-affected cell line will help facilitate investigation of the molecular basis of CWD prion pathogenesis and confirmation of CWD prion infectivity in vitro. KEY WORDS: CWD, infection, lentivirus, prion, RK13 cells. doi: 10.1292/jvms.12-0061; J. Vet. Med. Sci. 74(10): 1377-1380, 2012 Chronic wasting disease (CWD) is a neurodegenerative disorder in cervids and a member of the transmissible spongiform encephalopathies (TSEs), also known as prion diseases. TSEs are characterized by abnormal prion proteins (PrP Sc ) acting as infectious agents, which are generated by posttranslational modification of normal prion proteins (PrP C ) that accumulate in the brain and lead to diseases [10,13]. No TSEs have been found in any animal species in the Republic of Korea except for CWD, which is known to have originated from imported elk from Canada [12]. National CWD surveillance has been conducted since 2001, and subsequent cases were found in farmed elk in 2004 and 2005 [6]. More recently, CWD cases were detected in farmed elk, sika deer, red deer, and crossbred sika and red deer in 2010. This is the first report of natural CWD cases in red deer and sika deer (unpublished data). Epidemiologically speaking, it is possible that these indigenous CWD cases are associated with the imported CWD-affected elk.Several lines of persistently prion-affected cells have represented relevant and useful experimental models [14]. Mouse neuroblastoma cell lines (N2a) are the most commonly used; however, their use is limited by the fact that they are susceptible only to mouse-adapted prion strains. In contrast, rabbit kidney epithelial cells (RK13), which overexpress exogenous sheep PrP C , have been confirmed to be susceptible to natural sheep scrapie [3]. On the other hand, CWD prion persistently infected cell lines have been established from transformed primary mule deer brain cells [11]. Attempts to establish persistent CWD prion infection in a cell line have previously been unsuccessful, resulting in transient PrP Sc accumulation in elk PrP C -expressing RK13 cells [1].In this study, we attempted to establish a CWD prion persistently infected cell line to help characterize CWD prions, which are currently causing sporadic disease in South Korea. We tested the lentiviral protein expression system [5] as an approach method because it has broad tropism to a range of species and cell types, a stably expressed cell line is easily selected using safe laboratory ...