Uporaba umetnih transkripcijskih regulatorjev za uravnavanje prepisovanja genov v sesalskih celicah

magistrsko delo

Anže Verbič (Author), Darja Žgur-Bertok (Reviewer), Romana Marinšek-Logar (Mentor), Roman Jerala (Co-mentor)

Abstract

Uporaba načrtovanih DNA-vezavnih proteinov za natančen nadzor transkripcije poljubnih genov predstavlja enega izmed ključnih izzivov sintezne biologije. Do danes razviti pristopi temeljijo na genski fuziji efektorskih domen na načrtovane DNA-vezavne domene oziroma na kompetitivni vezavi načrtovanih DNA-vezavnih domen z endogenimi transkripcijskimi faktorji. V tej nalogi smo preučevali vpliv vezave načrtovanih TALE (ang. transcription activator-like effector) proteinov v bližino transkripcijskih aktivatorjev na osnovi cinkovih prstov Zif268 in Gal4. Ugotovili smo, da lahko z vezavo TALE proteinov v njuno bližino vplivamo na moč transkripcijske aktivacije, efekt pa je odvisen od pozicije in razdalje med vezavnimi mesti. TALE proteine lahko načrtujemo za vezavo v bližino endogenih transkripcijskih faktorjev, s čimer bi lahko nadzorovali izražanje endogenih genov celice.

Keywords

sintezna biologija;TAL efektorji;umetni transkripcijski faktorji;upravljanje prepisovanja genov;DNA vezavni proteini;cinkovi prsti;

Data

Language:	Slovenian
Year of publishing:	2017
Typology:	2.09 - Master's Thesis
Organization:	UL BF - Biotechnical Faculty
Publisher:	[A. Verbič]
UDC:	601.4:577.21
COBISS:	4857208
Views:	1135
Downloads:	615
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	English
Secondary title:	Use of artificial transcription regulators for transcriptional regulation in mammalian cells
Secondary abstract:	One of the main current areas of research in synthetic biology is the development of artificially designed DNA binding proteins for use in transcription regulation of endogenous genes. Currently used systems of artificial transcription regulation rely on mechanisms of binding site competition and on the use of effector domains for activation of repression of transcription. In this work, we examined the effects of proximal binding of TALE (transcription activator-like effector) proteins, to zinc-finger transcription factors Zif268 and Gal4. Binding of TALE proteins in the vicinity of Zif268 and Gal4 was found to have an effect on their transcriptional activation, dependent of the position and distance between binding sites. As TALE proteins can be designed to bind in the vicinity of endogenous transcription factors, this method could be employed as a novel approach to transcriptional regulation.
Secondary keywords:	synthetic biology;TAL effectors;artificial transkription factors;transcriptional regulation;DNA binding proteins;zinc fingers;
Type (COBISS):	Master's thesis/paper
Study programme:	0
Thesis comment:	Univ. v Ljubljani, Biotehniška fak., Študij mikrobiologije
Pages:	XI, 49 f., [5] f. pril.
ID:	10914768