Auxinak induzitutako degron (AID) sistemaren garapena funtzio ezezaguna duen proteina baten testuinguruan

Abstract

Auxinak induzitutako degron (AID) sistema proteinen kantitatearen murrizketa modu kontrolatuan lortzeko teknologia da, proteinen funtzioa aztertzeko aukera ematen duena. AID sistemari esker, mAID domeinua fusionatuta duten proteinak modu azkarrean degradatu daitezke, auxina molekula gehituz gero. Auxina degradazio sistemaren aktibatzaile gisa erabiltzen da eta sistemaren kontrola ahalbidetzen du. Lan honetan, AID sistema martxan jartzea lortu da TEDC2 genearen funtzioa ikertzeko asmoz. TEDC2 endogenoari mAID-mClover3 etiketa gehitu zaio CRISPR/Cas9 teknologia erabilita eta TEDC2-mAID-mClover3 fusio proteina eratu da. TEDC2-mAID-mClover3 zentrioloetan kokatzen dela frogatu dugu, eta auxina gehitzean proteina modu kontrolatuan degradatzea lortu da.Auxin induced degron (AID) system is a technology that allows the conditional depletion of a protein, being particulary useful for the functional analysis of uncharacterised proteins. In cells expressing the OsTIR1 plant protein, it is possible to degrade mAID-fused proteins with a very short half-time in the presence of auxin. In this work, we have successfully developed the AID system using TEDC2 gene as an example, thanks to its tagging with mAID-mClover3 by CRISPR/Cas9. In this way, TEDC2 has turned out to be located in the centrioles and, at the same time, it has been possible to achieve its depletion by the addition of auxin.