WO2018069782A3 - A combination of split orthogonal proteases with dimerization domains that allow for assembly - Google Patents

A combination of split orthogonal proteases with dimerization domains that allow for assembly Download PDF

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Publication number
WO2018069782A3
WO2018069782A3 PCT/IB2017/055902 IB2017055902W WO2018069782A3 WO 2018069782 A3 WO2018069782 A3 WO 2018069782A3 IB 2017055902 W IB2017055902 W IB 2017055902W WO 2018069782 A3 WO2018069782 A3 WO 2018069782A3
Authority
WO
WIPO (PCT)
Prior art keywords
proteases
split
orthogonal
dimerization domains
combination
Prior art date
Application number
PCT/IB2017/055902
Other languages
French (fr)
Other versions
WO2018069782A2 (en
Inventor
Roman Jerala
Mojca Bencina
Maja MESKO
Tina LEBAR
Jan LONZARIC
Tina FINK
Ziga STRMSEK
Fabio LAPENTA
Tjasa PLAPER
Katja LEBEN
Kosta CEROVIC
Estera MERLJAK
Nik FRANKO
Rok KRESE
Miha GRADISEK
Arne PRAZNIK
Nina JERALA
Lidija MAGDEVSKA
Samo ROSKAR
Ziga PUSNIK
Original Assignee
Kemijski Institut
En-Fist Center Odlicnosti
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Kemijski Institut, En-Fist Center Odlicnosti filed Critical Kemijski Institut
Priority to EP17794772.8A priority Critical patent/EP3526325A2/en
Publication of WO2018069782A2 publication Critical patent/WO2018069782A2/en
Publication of WO2018069782A3 publication Critical patent/WO2018069782A3/en

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/503—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from viruses
    • C12N9/506—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from viruses derived from RNA viruses

Abstract

Invention refers to combination of split orthogonal proteases that recognize and cleave target sequence of at least 6 amino acids. Parts of split orthogonal proteases are fused to dimerization domains that allow formation of the whole protease from two split protease parts. At least two orthogonal proteases are designed as split fragments fused to dimerization domains where dimerization can be induced with either light, chemicals or other input signal. Proteases cleave one or more target proteins that include cleavage site for one or more orthogonal proteases and act as a signal transducers, reporters or therapeutic proteins. With appropriately selected target proteins, logical circuits mediated by proteases can be prepared. The invention also relates to cells that contain expressed split proteases to transmit the signal.
PCT/IB2017/055902 2016-10-12 2017-09-27 A combination of split orthogonal proteases with dimerization domains that allow for assembly WO2018069782A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP17794772.8A EP3526325A2 (en) 2016-10-12 2017-09-27 A combination of split orthogonal proteases with dimerization domains that allow for assembly

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
SI201600252A SI25289A (en) 2016-10-12 2016-10-12 Combination of split orthogonal proteases with dimerization domains that enable the assembly
SIP-201600252 2016-10-12

Publications (2)

Publication Number Publication Date
WO2018069782A2 WO2018069782A2 (en) 2018-04-19
WO2018069782A3 true WO2018069782A3 (en) 2018-05-24

Family

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IB2017/055902 WO2018069782A2 (en) 2016-10-12 2017-09-27 A combination of split orthogonal proteases with dimerization domains that allow for assembly

Country Status (3)

Country Link
EP (1) EP3526325A2 (en)
SI (1) SI25289A (en)
WO (1) WO2018069782A2 (en)

Families Citing this family (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3701015B1 (en) * 2017-10-24 2022-04-06 Ulisse Biomed S.P.A. Amplification nanoswitch system based on split site-specific cleaving enzymes for the in vitro detection of target analytes and method for the detection of said target analytes
US10899823B2 (en) 2018-01-18 2021-01-26 California Institute Of Technology Programmable protein circuits in living cells
US11965191B2 (en) 2018-01-18 2024-04-23 California Institute Of Technology Programmable protein circuits in living cells
US11453893B2 (en) 2018-08-30 2022-09-27 California Institute Of Technology RNA-based delivery systems with levels of control
EP3844180A4 (en) 2018-08-31 2022-07-20 California Institute of Technology Synthetic protein circuits detecting signal transducer activity
WO2020146627A1 (en) 2019-01-10 2020-07-16 California Institute Of Technology A synthetic system for tunable thresholding of protein signals
EP3783104A1 (en) * 2019-08-20 2021-02-24 Kemijski Institut Coiled-coil mediated tethering of crispr-cas and exonucleases for enhanced genome editing
IT202000018064A1 (en) * 2020-07-27 2022-01-27 Univ Cattolica Del Sacro Cuore DEVELOPMENT OF A NEW ENGINEERED TOBACCO ETCH VIRUS (TEV) PROTEASE THAT CAN BE ACTIVATED IN THE CYTOSOL OR SECRETORY PATHWAY
CN112921053B (en) * 2021-02-02 2023-04-14 汕头大学 Dual-induction mCreER system capable of tracking cell differentiation and development and establishment and application thereof
CN114591442B (en) * 2022-03-01 2024-04-19 中国科学院深圳先进技术研究院 Light-regulated protease tool and matched substrate thereof
US20240011010A1 (en) * 2022-07-06 2024-01-11 California Institute Of Technology Synthetic protein-level neural network in mammalian cells
US20240124913A1 (en) * 2022-10-14 2024-04-18 California Institute Of Technology Protein-Based Signal Amplification

Citations (2)

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WO2014040129A1 (en) * 2012-09-12 2014-03-20 The University Of Queensland Protease-based biosensor
WO2015035452A1 (en) * 2013-09-12 2015-03-19 The University Of Queensland Bimolecular protease-based biosensor

Patent Citations (2)

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WO2014040129A1 (en) * 2012-09-12 2014-03-20 The University Of Queensland Protease-based biosensor
WO2015035452A1 (en) * 2013-09-12 2015-03-19 The University Of Queensland Bimolecular protease-based biosensor

Non-Patent Citations (10)

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Also Published As

Publication number Publication date
WO2018069782A2 (en) 2018-04-19
SI25289A (en) 2018-04-30
EP3526325A2 (en) 2019-08-21

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