Combination of split orthogonal proteases with dimerization domains that enable the assembly

• Main Authors: Arne Praznik, Maja Meško, Mojca Benčina, Tjaša Plaper, Lidija Magdevska, Roman Jerala, Jan Lonzarič, Katja Leben, Rok Krese, Nina Jerala, Estera Merljak, Miha Gradišek, Tina Fink, Tina Lebar, Samo Roškar, Fabio Lapenta, Kosta Cerović, Žiga Strmšek, Nik Franko, Žiga Pušnik
• Format: Patent
• Language: English; Slovenian
• Published: 30.04.2018
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

Izum se nanaša na kombinacijo ortogonalnih cepljenih proteaz, ki prepoznavajo tarčno zaporedje, ki obsega vsaj 6 aminokislin, povezanih z dimerizacijskimi domenami, ki omogočajo sestavljanje razcepljenih proteaz. Vsaj dve ortogonalni proteazi sta pripravljeni kot razcepljena fragmenta v povezavi z dimerizacijskimi domenami, kjer je dimerizacija lahko inducibilna s svetlobnim, kemičnim ali drugim vhodnim signalom. Proteaze cepijo enega ali več tarčnih proteinov, ki vsebujejo tarčno zaporedje za eno ali več ortogonalnih proteaz in delujejo kot nadaljnji posredniki signala, poročevalski proteini ali terapevtski proteini. Z ustrezno izbranimi tarčnimi proteini lahko pripravimo s proteazami posredovana logična vezja. Izum se nanaša tudi na celice, ki vsebujejo izražene razcepljene proteaze za prenos signala. 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.