Patent application EP19177622A (EP), filed 2019-05-31
Publication number: EP3744839A1
Title: A GENETICALLY ENCODED PROTEIN SYNTHESIS INHIBITOR (GEPSI)
Abstract: [0001]    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.
IPC codes: A61K  48/    00            A I
C07K  14/   415            A I
C12N  15/    64            A I
C12N   9/    24            A I
Claims: number: 1
text: An expression system for a genetically encoded protein synthesis inhibitor having RNA N-glycosidase activity comprising:
number: 2
text: The system of claim 1,
wherein the first component comprises an α-chain from a class 3 ribosome-inactivating protein (RIP), particularly an α-chain from a Panicoideae RIP, more particularly a maize RIP α-chain, or a functional variant thereof,
and/or
wherein the second component comprises a β-chain from a class 3 ribosome-inactivating protein (RIP), particularly a β-chain from a Panicoideae RIP, more particularly a maize RIP β-chain, or a functional variant thereof.
number: 3
text: The system of claim 1 or 2,
wherein the first nucleic acid sequence and the second nucleic acid sequence have different transcriptional starting points.
number: 4
text: The system of any one of claims 1-3,
wherein the first component comprises an amino acid sequence selected from:
number: 5
text: The system of any one of claims 1-7,
wherein the first expression control sequence and/or the second expression control sequence are inducible expression control sequences,
wherein the first and/or second expression control sequences may be inducible by temperature shift, by light, by addition or removal of one or more chemical substances (e.g. of synthetic or biological origin) or by any combination thereof, and
wherein the inducible expression control sequences may be chosen from tetracycline-controlled expression control sequences, hormone-controlled expression control sequences, light-controlled expression control sequences, and/or heat-shock expression control sequences.
number: 6
text: The system of any one of claims 1-5,
further comprising:
(d) a third nucleic acid sequence encoding a regulator, e.g. an activator for the first and/or second expression control sequence in operative linkage with a third expression control sequence.
number: 7
text: The system of claim 6,
wherein the third expression control sequence is a cell- and/or tissue-type specific expression control sequence,
wherein the third expression control sequence may be an expression control sequence specific for cells from brain tissue or specific for cells forming pre-cancerous or cancerous tissue.
number: 8
text: The system of any one of claims 1-7,
which is inducible by a site-specific recombinase, e.g. a Cre recombinase.
number: 9
text: The system of any one of claims 1-8,
which provides spatial and/or temporal expression control, particularly spatial and temporal expression control for the protein synthesis inhibitor.
number: 10
text: A host comprising an expression system of any one of claims 1-9, wherein the host may be an isolated cell, a cell preparation, a cell culture, an organoid, a tissue or an organ, particularly of mammalian origin, and more particularly of human origin, or a non-human organism, particularly of mammalian origin.
number: 11
text: The host of claim10,
wherein the expression system provides spatial and/or temporal expression control, particularly spatial and temporal expression control for the protein synthesis inhibitor in the host.
number: 12
text: A method of inhibiting protein synthesis in a host, comprising the steps:
number: 13
text: The method of claim 12,
wherein (i) expression of the functional protein synthesis inhibitor is under spatial and/or temporal control, particularly under spatial and temporal control,
wherein (ii) expression of the functional protein synthesis inhibitor is a cell- and/or tissue-specific expression,
wherein (iii) expression of the functional protein synthesis inhibitor is reversible, and/or
wherein (iv) expression of the functional protein synthesis inhibitor is under conditions providing a recovery of protein synthesis in the host.
number: 14
text: An expression system of any one of claims 1-9 or a host of claim 10 or 11 for use in medicine, or for use in a screening procedure.
number: 15
text: A kit for providing expression of a genetically encoded protein synthesis inhibitor having N-glycosidase activity in a host, comprising:
Office action: search_report, dated 2020-12-02
Rejection reasons: novelty_epc_54
inventive_step_epc_56
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ref_id: EP0652231
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severity: novelty_destroying
ref_id: WO2012038950
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severity: background
ref_id: - TAN YUNG-CHIE ET AL, "Cloning and Expression of Oil Palm (Elaeis guineensisJacq.) Type 2 Ribosome Inactivating Protein inEscherichia coli", INTERNATIONAL JOURNAL OF PEPTIDE RESEARCH AND THERAPEUTICS, SPRINGER-VERLAG, DORDRECHT, NL, vol. 22, no. 1, doi:10.1007/S10989-015-9485-5, ISSN 1573-3149, (20150813), pages 37 - 44, (20150813), XP035922198 [X] 1-5,9-11,15 * the whole document * * materials and methods; pages  39,40 *
ref_type: npl
severity: novelty_destroying
ref_id: - FRANK SCHNÜTGEN ET AL, "A directional strategy for monitoring Cre-mediated recombination at the cellular level in the mouse", NATURE BIOTECHNOLOGY, (20030331), vol. 21, no. 5, doi:10.1038/nbt811, ISSN 1087-0156, pages 562 - 565, XP055108504 [YD] 8 * the whole document *
ref_type: npl
severity: obviousness
ref_id: - S. K.-Y. LAW ET AL, "A switch-on mechanism to activate maize ribosome-inactivating protein for targeting HIV-infected cells", NUCLEIC ACIDS RESEARCH, (20100617), vol. 38, no. 19, doi:10.1093/nar/gkq551, ISSN 0305-1048, pages 6803 - 6812, XP055015165 [A] 1-15 * the whole document *
ref_type: npl
severity: background
ref_id: - AKKOUH OUAFAE ET AL, "Biological activities of ribosome-inactivating proteins and their possible applications as antimicrobial, anticancer, and anti-pest agents and in neuroscience research", APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, SPRINGER BERLIN HEIDELBERG, BERLIN/HEIDELBERG, vol. 99, no. 23, doi:10.1007/S00253-015-6941-2, ISSN 0175-7598, (20150922), pages 9847 - 9863, (20150922), XP035870290 [A] 1-15 * the whole document *
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severity: background
Final disposition: pending (2020-12-02)