Preusmjeravanje adenovirusa tipa 5 na integrin α4β1

master's thesis

• Title (english): Retargeting adenovirus type 5 to integrin α4β1
• Author: Tihana Jelušić
• Mentor: Andreja Ambriović Ristov (mentor)
• Committee member: Andreja Ambriović Ristov (predsjednik povjerenstva)
• Committee member: Dubravka Hranilović (član povjerenstva)
• Committee member: Mladen Krajačić (član povjerenstva)
• Committee member: Maja Matulić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2009, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Gensko preusmjeravanje Adenovirusa tipa 5 (Ad5) ugradnjom sljedova u vlakno bi trebalo omogućiti ciljani unos gena. Cilj je ovog istraživanja konstruirati replikacijski defektan adenovirusni vektor uvođenjem ciljnog liganda CPLDIDFYC za integrin α4β1 u HI petlju glave vlakna, kako bi se preusmjerio Ad5 na integrin α4β1 (poznat i pod imenom VLA-4, engl. very late antigen). Rekombinantni Ad5VLA-4 virus konstruiran je manipulacijom cjelovitog Ad5 genoma kao plazmida u E. coli, pri čemu je korištena homologna rekombinacija u bakterijama. Ad5VLA-4 i kontrolni virus Ad5wt bez ciljnog slijeda, umnoženi su u kulturi bubrežnih stanica ljudskog embrija 293 i pročišćeni u gradijentu cezijeva klorida. Koristeći model ljudskih stanica rabdomiosarkoma, koje eksprimiraju integrin α4β1, pokazali smo povećanu djelotvornost transdukcije s Ad5VLA-4 u usporedbi s Ad5wt. Prema tome Ad5VLA-4 bi se mogao koristiti za gensku terapiju tumora koji eksprimiraju integrin α4β1.

Abstract (english)

Genetic retargeting of Adenovirus type 5 (Ad5) through insertion of sequences in the fiber protein should permit targeted gene delivery. The aim of this research is to construct replication defective adenoviral vector by incorporating α4β1 integrin-targeting ligand CPLDIDFYC into the HI-loop of the fiber protein, in order to retarget Ad5 to integrin α4β1 (also known as very late antigen 4, VLA-4). Construction of Ad5VLA-4 was performed by manipulation of the full length Ad5 genome as a stable plasmid in E. coli, using the bacterial homologous recombination machinery. Ad5VLA-4 and a control virus Ad5wt lacking targeting sequence, were multiplied in human embryonic kidney cell line-293 and purified by bending in CsCl gradients. On the rhabdomyosarcoma (RD) cell line, which express α4β1 integrin, Ad5VLA-4 exhibited increased transduction efficacy in comparison to Ad5wt. Therefore, Ad5VLA-4 could be suitable vector for gene therapy of tumors expressing α4β1 integrin.

• Keywords: preusmjeravanje; adenovirus; integrin α4β1; VLA-4
• Keywords (english): Ad retargeting; integrin α4β1; VLA-4
• Language: croatian
• URN:NBN: urn:nbn:hr:217:228201
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2018-03-12 11:11:36