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Načrtovanje hitro odzivnih poti za sesalske celice
Jovanovski, Maša (Author), Jerala, Roman (Mentor) More about this mentor... This link opens in a new window

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Abstract
Sintetična biološka logična vezja, podobno kot električna, zaznavajo vstopne informacije, jih predelajo ter na podlagi le-teh tvorijo specifične celične odzive. Večina do sedaj ustvarjenih logičnih vezij temelji na regulaciji transkripcije, ki je časovno potratnejša kot prenos signala preko proteinskih interakcij. Za zdravljenje določenih bolezni, kot je diabetes, je regulacija transkripcije prepočasna, saj za razvoj polnega signala potrebuje nekaj ur. V diplomski nalogi predstavljamo umetno ustvarjeno hitro odzivno signalno pot, ki je modularna ter med aplikacijami lahko prenosljiva. V HEK293T celice smo vnesli na obvitih vijačnicah ter proteolizi osnovanih konstruktih temelječ sistem za logično procesiranje signala. Sistem je inducibilen z rapamicinom ter abscizinsko kislino, vendar bi se ga dalo prilagoditi, da se odziva na eksogene signale kot sta ultrazvok ter svetloba, ali na endogene signale. Testirali smo logično funkcijo IN, preko katere smo le ob vnosu obeh izbranih vstopnih signalov hkrati dobili željen signal že 10 minut po stimulaciji celic. S prilagoditvijo cepitvenih mest, lahko s takšnim sistemom izvajamo tudi ostale Boolove logične funkcije, kar ustvarjenemu sistemu omogoča še večjo uporabnost na različnih znanstvenih področjih.

Language:Slovenian
Keywords:logična vezja, proteoliza, biotehnologija, sintezna biologija, obvite vijačnice
Work type:Bachelor thesis/paper (mb11)
Tipology:2.11 - Undergraduate Thesis
Organization:BF - Biotechnical Faculty
Year:2019
Publisher:[M. Jovanovski]
UDC:602.64:601.4:577.21(043.2)
COBISS.SI-ID:9258105 Link is opened in a new window
Views:68
Downloads:27
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Secondary language

Language:English
Title:Design of fast responsive signaling pathways in mammalian cells
Abstract:
Much like electrical circuits, synthetic biological logic circuits have the ability to sense inputs, process them and form celullar responses based on inputs. So far, most logic circuits are based on regulating transcription, where signal transduction is more time consuming than signal transduction through protein-protein interactions. For treating certain illnesses like diabetes, transcription regulation is too slow, as signals are fully developed after a few hours. This Bachelor thesis presents a synthetical fast responsive signalling pathway, which is highly modular and thus very easily transferable between different applications. We have tranfected HEK293T cells with constructs based on coiled coils and proteolysis for logical processing of signals. The system is inducible with rapamycin and abscisic acid but it can be modified to cell reaction to other exogenous signals like ultrasound and light or to cellular endogenous signals. We have demonstrated the Boolean function AND, through which the desired signal was obtained 10 minutes after stimulating cells only with both input signals. By adjusting the cleavage sites, the system is able to execute other Boolean functions as well, thus ensuring the system an even greater possibility of interdisciplinary use.

Keywords:logic circuits, proteolysis, biotechnology, synthetic biology, coiled coils

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