Cell lines for virus production and methods of use
AssigneesUniversity of Georgia Research Foundation, Inc., The Government of the USA as represented by the Secretary of the Dept. of Health and Human Service, Thermo Fisher Scientific Biosciences Inc.
InventorsJon Michael Karpilow, Mark Steven Oberste, Ralph A. Tripp, Stephen M. Tompkins
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Abstract
Provided herein are engineered cell lines. In some embodiments, cells of an engineered cell line have altered expression of a gene and/or altered expression of an miRNA, wherein the altered expression results in increased or decreased production of a virus. The virus is a picomavirus, such as a poliovirus or Enterovirus 71. Also provided herein are methods for using the engineered cells to produce virus, and methods for treating a subject having or at risk of having a viral infection.
Claims39
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An engineered cell line, wherein cells of the engineered cell line comprise decreased expression of at least one coding region selected from Table I compared to a control cell line, and optionally wherein the at least one coding region is selected from ZNF205, CNTD2, SEC61G, ETS1, TAF1L, MCCD1, LY6G6C, BTN2A1, GLXP3, GCGR, or EP300.
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An engineered cell line, wherein cells of the engineered cell line comprise increased expression of at least one coding region selected from Table II compared to a control cell line.
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An engineered cell line, wherein cells of the engineered cell line comprise increased expression of at least one miRNA selected from miR-520e, miR-1256, miR-520d-3p, miR-513a-5p, miR-519c-3p, miR-1270-2, miR-3187, miR-5763p, miR-22, miR-520c-3p, and miR-9 compared to a control cell line.
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An engineered cell line, wherein cells of the engineered cell line comprise decreased expression of at least one endogenous miRNA selected from Table IV compared to a control cell line.
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A method for producing a virus comprising:
- providing an engineered cell line wherein cells of the cell line comprise a virus, wherein cells of the engineered cell line comprise
- (i) decreased expression of at least one coding region selected from Table I compared to a control cell line,
- (ii) increased expression of at least one coding region selected from Table II compared to a control cell line,
- (iii) increased expression of at least one miRNA selected from miR-520e, miR-1256, miR-520d-3p, miR-513a-5p, miR-519c-3p, miR-1270-2, miR-3187, miR-5763p, miR-22, miR-520c-3p, and miR-9 compared to a control cell line, or
- (iv) decreased expression of at least one endogenous miRNA selected from Table IV compared to a control cell line;
- incubating the engineered cell line under conditions suitable for the production of the virus by the cells;
- and
- harvesting the virus produced by the cells.
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A method of treating a subject having or at risk of having a viral infection comprising increasing, in cells of the subject, expression of a coding region selected from Table I, (i) inhibiting, in cells of the subject, expression of a coding region selected from Table II, (ii) inhibiting expression, in cells of the subject, of an endogenous miRNA selected from miR-520e, miR-1256, miR-520d-3p, miR-513a-5p, miR-519c-3p, miR-1270-2, miR-3187, miR-5763p, miR-22, miR-520c-3p, and miR-9, or (iv) increasing, in cells of the subject, expression of an miRNA selected from Table IV.
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Dependencies are inferred from the claim text.