Compositions and methods for enhancing gene expression
AssigneeJanssen Pharmaceuticals, Inc.
InventorsKurt Iver Kamrud, Maung Nyan Win, Nathaniel Stephen Wang, Jason L. DeHart
Life of this patent
Estimating the term…
Abstract
The present disclosure generally relates to nucleic acid molecules for use in regulating gene expression. Disclosed herein include nucleic acid molecules containing one or more structural elements of the viral capsid enhancer operably linked to a coding sequence of a gene of interest. In some embodiments, the viral capsid enhancer comprises a Downstream Loop (DLP) from a viral capsid protein, or a variant of the DLP.
Claims76
A nucleic acid molecule, comprising:
- a first nucleic acid sequence encoding one or more RNA stem-loops of a viral capsid enhancer or a variant thereof;
- and
- a second nucleic acid sequence operably linked to the first nucleic acid sequence, wherein the second nucleic acid sequence comprises a coding sequence for a gene of interest (GOI).
-5. (canceled)
(canceled)
-12. (canceled)
(canceled)
-18. (canceled)
(canceled)
-26. (canceled)
A method for producing a polypeptide of interest in a cell, comprising introducing a RNA molecule into the cell, wherein the RNA molecule comprises one or more RNA stem-loops of a viral capsid enhancer or a variant thereof, and a coding sequence for the polypeptide of interest, thereby producing the polypeptide of interest in the cell.
(canceled)
-36. (canceled)
(canceled)
A method for producing a messenger RNA (mRNA) in a cell, comprising introducing into the cell a nucleic acid molecule comprising a first nucleic acid sequence encoding one or more RNA stem-loops of a viral capsid enhancer or a variant thereof, and a second nucleic acid sequence operably linked to the first nucleic acid sequence, wherein the second nucleic acid sequence comprises a coding sequence for a gene of interest (GOI), thereby producing a mRNA of the GOI.
-58. (canceled)
-68. (canceled)
A nucleic acid molecule comprising a nucleic acid sequence encoding a modified viral RNA replicon, wherein the modified viral RNA replicon comprises:
- a first nucleic acid sequence encoding one or more structural elements of a viral capsid enhancer or a variant thereof, wherein the viral capsid enhancer is derived from a first virus species, and
- a second nucleic acid sequence derived from a second virus species encoding at least one nonstructural viral protein or a portion thereof,
- wherein the first nucleic acid sequence is operably linked upstream to the second nucleic acid sequence.
-73. (canceled)
(canceled)
-82. (canceled)
-87. (canceled)
(canceled)
-100. (canceled)
A nucleic acid molecule comprising a nucleic acid sequence encoding a modified non-alphavirus RNA replicon, wherein the modified non-alphavirus RNA replicon comprises a first nucleic acid sequence encoding one or more structural elements of a viral capsid enhancer or a variant thereof.
-113. (canceled)
-117. (canceled)
A method for conferring a resistance to the innate immune system in a subject, comprising administering to the subject a nucleic acid molecule comprising a nucleic acid sequence which encodes a modified viral RNA replicon, wherein the modified viral RNA replicon comprises:
- a first nucleic acid sequence encoding one or more structural elements of a viral capsid enhancer or a variant thereof, wherein the viral capsid enhancer is derived from a first virus species, and
- a second nucleic acid sequence derived from a second virus species encoding at least one nonstructural protein or a portion thereof,
- wherein the first nucleic acid sequence is operably linked upstream to the second nucleic acid sequence, and wherein expression of the modified replicon RNA encoded by the nucleic acid molecule confers a resistance to innate immune response in the subject.
A method for producing a polypeptide of interest in a subject, comprising administering to the subject a nucleic acid molecule comprising a nucleic acid sequence which encodes a modified viral RNA replicon, wherein the modified viral RNA replicon comprises:
- a first nucleic acid sequence encoding one or more structural elements of a viral capsid enhancer or a variant thereof, wherein the viral capsid enhancer is derived from a first virus species, and
- a second nucleic acid sequence derived from a second virus species encoding at least one nonstructural protein or a portion thereof,
- wherein the first nucleic acid sequence is operably linked upstream to the second nucleic acid sequence.
(canceled)
-127. (canceled)
(canceled)
-133. (canceled)
-136. (canceled)
(canceled)
-153. (canceled)
A method for conferring a resistance to the innate immune system in a subject, comprising administering to the subject a nucleic acid molecule comprising a nucleic acid sequence encoding a modified non-alphavirus RNA replicon, wherein the modified non-alphavirus RNA replicon comprises a first nucleic acid sequence encoding one or more structural elements of an alphavirus capsid enhancer and wherein expression of the modified non-alphavirus RNA replicon encoded by the nucleic acid molecule confers a resistance to innate immune response in the subject.
A method for producing a polypeptide of interest in a subject, comprising administering to the subject a nucleic acid molecule comprising a nucleic acid sequence encoding a modified non-alphavirus RNA replicon, wherein the modified non-alphavirus RNA replicon comprises a first nucleic acid sequence encoding one or more structural elements of an alphavirus capsid enhancer.
A method for producing a polypeptide of interest, comprising culturing a host cell comprising a nucleic acid molecule which comprises a nucleic acid sequence encoding a modified non-alphavirus RNA replicon, wherein the modified non-alphavirus RNA replicon comprises a first nucleic acid sequence encoding one or more structural elements of an alphavirus capsid enhancer.
-170. (canceled)
-174. (canceled)
Dependencies are inferred from the claim text.