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US 9,359,595 B2Granted
Method of preparing cells susceptible to transmissible spongiform encephalopathy and the creation of TSE persistently infected cells
AssigneeAnimal, Plant and Fisheries Quarantine and Inspection Agency
InventorsDong-Seob Tark, Hyo-Jin Kim, Hyun-Joo Shon, Yoon-Hee Lee, Min-Jeong Kim, Eun-Im Yun, In-Soo Cho, Chang-Hee Kweon, Yi-Seok Joo, Otto Windl, Michael Neale
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Abstract
Disclosed is a method for preparation of transmissible spongiform encephalopathy (TSE) susceptible cells, and such cells may be efficiently used in diagnosis of TSE, etiology studies and development of novel therapeutics, etc.
Claims2
A method for preparation of a cell persistently infected with bovine spongiform encephalopathy (BSE), comprising:
- extracting a DNA comprising a normal prion protein gene from bovine brain and cloning the normal prion protein gene into a lentivirus transfer vector;
- inserting the lentivirus transfer vector comprising the normal prion protein gene into a lentivirus to produce a recombinant lentivirus;
- inoculating a MDBK cell line with the recombinant lentivirus to produce a transduced cell expressing a normal prion protein;
- selecting one of multiple transduced cells that expresses the normal prion protein;
- inoculating the selected transduced cell expressing the normal prion protein with a BSE-infected bovine brain homogenate
- to produce a clone capable of exhibiting infection even after 26 passages by continuous subculture.
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