Genetically encoded protein synthesis inhibitor
AssigneeMax-Planck-Gesellschaft zur Forderung Wissenschaften e.V.
InventorsErin M. Schuman, Maximilian Heumüller, Caspar Glock
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Abstract
The present invention relates to an expression system for a genetically encoded protein synthesis inhibitor containing RNA N-glycosidase activity split into two components. The expression system can be combined with genetic targeting systems to achieve cell- and/or tissue-type-specific and/or temporally-specific control of protein synthesis in a host, particularly in a mammalian host.
Claims17
An expression system for a genetically encoded protein synthesis inhibitor having RNA N-glycosidase activity comprising:
- (a) a first nucleic acid sequence encoding a first component of the protein synthesis inhibitor in operative linkage with a first expression control sequence, wherein the first component comprises an amino acid sequence as shown in SEQ ID NO:1 or an amino acid sequence having an identity of at least 95% thereto;
- and
- (b) a second nucleic acid sequence encoding a second component of the protein synthesis inhibitor in operative linkage with a second expression control sequence, wherein the second component comprises an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence having an identity of at least 95% thereto;
- and
- (c) a third nucleic acid sequence encoding a trans-acting activator for the first and second expression control sequences in operative linkage with a third expression control sequence,
- wherein the first expression control sequence or the second expression control sequence comprise an inducible expression control sequence responsive to an external stimulus, wherein the external stimulus is external to the expression system, thereby providing the expression system with reversible inhibition of protein synthesis and temporal control,
- wherein the third expression control sequence is a cell- and/or tissue-type specific expression control sequence,
- wherein the expression system is adapted for expressing the first nucleic acid sequence and the second nucleic acid sequence separate from each other,
- wherein the first component and the second component together form a complex having RNA N-glycosidase activity, and
- wherein the first component alone and the second component alone lack RNA N-glycosidase activity.
A kit for providing expression of a genetically encoded protein synthesis inhibitor having RNA N-glycosidase activity in a host, comprising:
- (a) a first nucleic acid sequence encoding a first component of the protein synthesis inhibitor in operative linkage with a first inducible expression control sequence, wherein the first component comprises an amino acid sequence as shown in SEQ ID NO:
- 1 or an amino acid sequence having an identity of at least 95% thereto;
- and
- (b) a second nucleic acid sequence encoding a second component of the protein synthesis inhibitor in operative linkage with a second inducible expression control sequence, wherein the second component comprises an amino acid sequence as shown in SEQ ID NO:2 or an amino acid sequence having an identity of at least 95% thereto;
- wherein the first nucleic acid sequence and the second nucleic acid sequence are provided for expression separate from each other,
- wherein the first inducible expression control sequence or the second inducible expression control sequence is responsive to an external stimulus, wherein the external stimulus is external to the host, thereby providing the host with reversible inhibition of protein synthesis,
- wherein the first component and the second component together form a complex having RNA N-glycosidase activity, and
- wherein the first component alone and the second component alone lack RNA N-glycosidase activity, and
- (c) a cell- and/or tissue-type specific expression control sequence in operative linkage with a third nucleic acid sequence encoding a trans-acting activator responsible for restricting expression of (a) and (b) to a targeted cell and/or tissue.
Dependencies are inferred from the claim text.